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295 Cards in this Set
- Front
- Back
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cDna is made because bacteria cannot go through
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RNA processing and cut out irrelevant parts
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PCR is the same as
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thermo-cycler
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Thermo aquaticus is an _________ bacteria
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archean
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PCR goes through _____ cycles and end up with _______ molecules
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30; over a billion (2^30)
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_______ is a major opponent to agricultural biotechnology
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Jeremy Rifkin
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_____ corn is used for animals only
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Starlink
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Classical breeding does what?
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mutate within same species
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_____ virus kills papayas
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Ringspot
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An example of an organic pesticide is
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BT
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PRoblem with pesticides is
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target becomes resistant to pesticide
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A refuge is
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a place where nonresistant pests can live
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_______ was group that fired at MSU and tried to stop GMO's
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Earth Liberation
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________ engineered GM salmon
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Aqua Bounty Farms
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Blue pipet min and max
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100 micro-1000micro
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Yellow pipet max and min
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20 micro- 200 micro
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White pipet max and min
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.5 micro- 10 micro
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Chelex are
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plastic beads that attract heavy metals
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______ separates DNA while ____ puts DNA primers to target
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95'C; 60'C
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Order of materials in blotting step of SOuthern Blotting
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alkaline solution, sponge, gel, nitrocellulose paper, paper towels
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The purpose of the alkaline solution in S. blotting is to
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pull sample against filter and slightly denature
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"Ready-to-go" beads are
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little while balls that have taq polymerase and nucleotides ready to go
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The primary method of analyzing cloned DNA is
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gel electrophoresis
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Agar means
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seaweed
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Agarose gel separates
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DNA and some proteins
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Agarose gel is usually set up
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horizontally
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You use _______ agarose if you want to separate fragments that are small in size
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concetrated
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NEgative DNA will go to the
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anode (positive) side
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Shorter fragments go _____ in a gel electrophoresis
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farther
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Chemical dyes are 3-ringed structures that look like what?
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A step in the DNA ladder
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Dyes become _______ in DNA
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intercolated
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An electrophoresis is run in a _____ solution
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buffer
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The buffer solution in the gel electrophoresis is used for what?
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Keeping the pH constant
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The buffer solution in a gel electrophoresis contains two substances. WHat are they?
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A weak base and its salt
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The weak base in the buffer is usually
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Tris
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The complementary salt to Tris is
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boric acid
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together, the usual buffer is called
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Tbe buffer (Tris)
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If there is no buffer in the electrophoresis, then what will happen?
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The cathode end will become acidic and the anode will become alkaline
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______ for longer times will separate DNA better than ______ all at once
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low voltage; high voltage
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DNA will show up after the gel has been
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stained
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The usual stains (under UV light) are
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Methylene Blue and Ethidium Bromide
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Which stain works better?
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Ethidium Bromide
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What gel has a tighter mesh of polymers than agarose gel and can distinguish between one base pair length?
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Polyacrylamide gels
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One base pair length is
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.34 nm
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Polyacrylamide gels must be run
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horizontally
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______can be used to tell if a gene is herterozygous or homozygous or if there are restriction sites witihin the gene
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Gels
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For sickle cell anemia, a normal cell has ____ band, sickle cell has ____, and heterozygous has ____ bands
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1, 2, 3
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A Southern Blotting Technique is a key technique is
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DNA identification
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Northern Blots are used for
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RNA
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Western Blots are used for
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polypeptide chains (denatured proteins)
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DNA fingerprinting was created by
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SIr Alec Jeffreys
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______regions of the DNA are many time more polymorphic than coding regions
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Noncoding
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Why are noncoding regions of DNA more polymorphic than coding regions?
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They are not highly conserved
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Sets of known regions that are highly polymorphic and bordered by certain restriction sites are known as
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Short tandem repeats (STR)
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STR stands for
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short tandem repeat
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MAtching sets of ____ are used identification markers in forensic
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STR's
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All forensics labs use same __________ and calculate percentage of population that hold those genes.
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13 tandem repeats
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An autoradiograph is an
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x-ray film in SOuthern Blotting
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The purpose of Southern Blotting is to
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determine if organisms contain similar DNA
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In Southern Blotting, you compare two organisms by
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looking for a specific nucleotide sequence and comparing the size of the fragments
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RFLP is a
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restriction fragment length polymorphism
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What is a RFLP?
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Take a piece of DNA, use restriction enzymes to make little fragments
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The restriction enzyme used in our lab was
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HAE3
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The gene from our lab was for
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tasting bitterness
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_____percent of DNA is STR
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50
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In order to mapping to work, an SNP must be physically close to
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the allele
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The closer the SNP is to the allele, the higher the
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degree of certainty
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SNP change
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restriction sites
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The Sanger method for sequencing does what?
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Takes PCRed DNA, divide into four samples, add a primer that is complementary to 3', add unique ddnucleotide, run gel
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Dideoxy nucleotide does not have
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3 OH
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Each sample in the Sanger method only gets
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one dideoxy nucleotide
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Restriction Fragment analysis is
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looking at length of DNA and comparing the two diff DNA molecules
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Polymorphisms mean
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many forms
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Polymorphisms are
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variations in DNA sequence among population
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Dideoxyribonucloetide chain termination method is
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used to find sequence of DNA
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A type of Northern Blotting is
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Reverse Transcriptase Polymerase Chain Reaction (RT-PCR)
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RT-PCR is when
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mRNA is made into cDNA
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What is process of RT-PCR?
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cDNA amplified by PCR using primers specific to a desirable gene, electrophoresis will reveal amplified DNA products with desired genes
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in situ hybridization is what?
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intact organism is used to determine which cells are expressing certain genes, probes used with mRNA of embryo
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_____allows genome wide expression study
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DNA microarray assay
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What does DNA Microarray do?
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Take diff DNA, put into different spots with a complement, see where genes are
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RNA interference is known as
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RNAi
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RNAi is the
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method of silencing expression of selected genes
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How does RNAi work?
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Takes synthetic double stranded RNA and triggers breakdown or blockage of corresponding mRNA
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SNP is where variation is found in at least _______ of the population
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1%
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Organismal cloning
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one or more organisms genetically identical to the parent
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Genomic equivalence
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if all cells of an organims have same genes
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Totipotent cells are ones that
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can mature and dedifferentiate and then give rise to all specialized cells
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Nuclear transplantation is when one
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removes the nucleus of un/fertilized cells and replaces it with differentiated cells
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Reproductive Cloning is the
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production of new individuals
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Stem cell is an
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unspecialized cell that can reproduce indefinitely and differentiate into specialized cells of one or more types
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Embryonic Stem (ES) cells
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reproduce indefinitely and differentiate into many diff types
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Adult stem cells
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can only give rise to multiple types of cells
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pluripotent cells
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capable of differentiating into many different cell types
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induced pluripotet stem (iPS) cells are
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differentiated cels that are turned into ESS cells by using retroviruses
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iPS cells are used to overcome
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nonfunctional tissue
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In Microarray, you put what in each well?
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single stranded cDNA
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______is the most common vector
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Ti plasmid
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JellyFish used in our experiment was
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Aequorea Victoria
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Plants have been made to last longer using
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antisense RNA
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tPA is
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tissue plasminogen activator
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Along with herbicides, plants have become more _______ resistant
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insect
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Antisense RNA prevents What?
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Ripening and Spoilage
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Biotechnology has enlarged the ______ parts of plants
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economical
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Vitamin integration is
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improving the nutrition of plants
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People have allergic reaction to _____, which is what is changed in biotechnology
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proteins
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Golden Rice is what?
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Rice that has the gene for beta carotene
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________ children in the developing world go blind each year form vitamin A deficiency
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250,000-300,000
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_______ of blinded children die within a year
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50%
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Rhizobiums are the ____ most important organism in the world
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3rd
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If Rhizobium job is integrated into the gene of plants, then what happens?
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plants can do nitrogen fixation themselves and the world food shortage is averted
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Microorganism can extract______ from environment and transform them into harmless or usable forms
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heavy metals
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Microorganisms ca degrade ____ that cannot be easily degraded
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organic compounds
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Microorganisms may be able to clean up ____ spills and _____ drops
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oil; mercury
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New bacteria can handle radioactive compounds up to _____ higher than humans
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1000s of times
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Bacteria can take uranium metals and turn them into_____ form. Why is this beneficial?
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Nonwater; it is harder to get into the water supply and drops the exposure risks
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______ can be used for myelogenous leukemia
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Gleevic
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Transplant organs must be _______ compatible
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MHC
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MHC is
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major histone complex
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Transgenic creatures are creatures that are
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developed from two diff species/genes
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______ are prime candidates for gene fixing
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bone marrow stem cells
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The best job in medicine has been improving _____ cells
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immune
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SCID stands for
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severe combined immuno deficiency
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Problem with CF transformation is
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cells reject new DNA; vector problem
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CF main symptom is ____; main problem is _____
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thick mucus in lungs; chloride ion channel does not work properly
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A vector is
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anything that carries genetic material
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Transduction is
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transformation exclusively for viral vectors
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The most common genetic problem for caucasians is
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Cystic Fibrosis
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Control mechanisms are
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promoter sequences
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For pharmaceuticals, one can insert genes into ____ to make high quantities of human _____
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bacteria; protein
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A stroke is a
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clot or rupture in the blood vessels
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BActeria use ___, ____, and _____ to defend against phages
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RE, methylated adenine and cytosine
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What does tPA do?
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Dissolves blood clots and reduces risk of heart attacks
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Gene cloning is useful for what two reason?
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1. Harvest protein product
2.Make gene copies (library development) |
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Nucleases allow what?
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reproducible fragments to be made
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When is tPA effective?
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up to 3 hours after a stroke
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Restriction enzymes were found in late
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1960s
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Nucleases go both ways, or are
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pallindromic
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Re cut sequences about _____ base pairs long
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3-8
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Some RE cut ____ the DNA and others make sticky ends
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straight through
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Every RE has a specific
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restriction site
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In order to go through cloning, you need a good
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cloning vector
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RE come in two flavors: _______ & _______
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endonucleases and ectonucleases
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RE needs to work with ____ and _____
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the site; DNA fragment
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A good cloning vector needs to be
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1.antibiotic resistant
2.have a marker gene |
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Gene library must be house in
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a specific medium or organism
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Gene library is
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collection of bunches of DNA
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What is a marker gene?
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Gene that makes a visual difference in colonies when disrupted
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Transformation is the step of
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putting a plasmid back into a bacteria
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Broken marker gene will result in
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same color colonies
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PLasmid that got the foreign DNA is called
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recombinant plasmid
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Genetic recombination is the
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combination of DNA from two sources
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Working marker gene results in
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different color colonies
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An example of a marker gene is
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lacZ
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Transformation, tranduction, and conjugation are all examples of
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genetic recombination
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The plasmid that's going to get foreign DNA is the
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cloning vector
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An example of a genomic library is
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when a plasmid has become recombinant
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Horizontal gene transfer is the
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movement of genes from members of different species`
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Actual definition of transformation is
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genotype of a prokaryotic cells is altered by uptake of foreign DNA
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transduction definition
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phages carry one host cell to another
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R. Plasmids are
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plasmids that carry resistance genes
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What allows for study of particular cell function and gene expression?
|
making cDNA
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Hfr cells are known as
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high frequency of recombination
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_____ of caucasians are carrier of CF gene
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1/25
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cDNA is made how?
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mRNA has complete DNA made, broken down, DNA gets compliment
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An Hfr cells is one that
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has an F factor on its chromosome
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The Solution that is sloshed with filter paper has
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a probe with gene of interest on it
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What is the human germline gene therapy?
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making genetic changes that will be inherited by future generations
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mEtabolomics is the
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study of cDNA and metabolic processes
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Who was the first to put an artificial chromosome in an animal and what animal was it?
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Chromos; a mouse
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Dna is usually injected via
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microinjection
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Reverse transcriptase is used in what two processes?
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HIV, cDNA
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How are artificial chromosome inherited?
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Like a natural chromosome
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Microinjection _____ with DNA because it's ______
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doesn't; too large
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Reverse transcriptase does what?
|
backwards transcription
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How do you start making artificial chromosomes?
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use DNA manufacturing enzymes to duplicate stubby arms and extend them with satellite DNA
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Random genes can be passed on, but with no
|
therapeutic effect
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Child born with HIV is so because of
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bodily fluid transfer
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In order to be effective, a plasmid must have it's own promoter sequence, or
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bacterial promoter sequence
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An expression vector is
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a plasmid that has a powerful bacterial promoter sequence
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Yeasts are _____ and thus can go through RNA processing
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eukaryotic
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Inserting chromosomes creates
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a new species
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Bacterial promoter sequence is used for
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high efficiency transcription
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Yeasts have ______ & ______ as well
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endoplasmic reticulum; plasmid
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Artificial chromosome behaves as a
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real chromosome
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_____ are closes Eukaryotic relative to bacteria
|
yeast
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Hybrid plasmids have been created using
|
yeast and bacterium
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What's wrong with germ line therapy?
|
You don't have the right to modify future generations
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cDNa can show the ____ that are found in any tissue
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genes
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Only ____ amount of gene is expressed with each cell
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small
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_____ techniques of competency
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2
|
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An example of a 2+ ion salt is
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CaCl(2)
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"making cells competent" or "increasing competency" means
|
making transformation more effective
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One method of competency is _____, followed by a ______
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2+ ion salt water bath; heat shock
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Another technique of competency is
|
electroporation
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Bacteria cells must be made _______ to accept DNA readily
|
competent
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The 2+ ion salt water bath and heat shock allow ____ to form
|
pores
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What is electroporation?
|
Electric charge used with Eukaryotic too
|
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One can use a ______ as a vector to send DNA into bacteria
|
bacteriophage
|
|
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For plant cells, one can use a _____, coat pellets with ____, and shoot the pellets
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baby shot gun; DNA
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BAC's are
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bacterial artificial chromosomes
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BAC can carry DNA insert of length
|
100-300 kilobase pairs
|
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Someone can also ____ DNa into cells directly
|
inject
|
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What is a bacterial artificial chromosome?
|
type of vector used in library construction- large plasmids trimmed down so they just contain genes necessary for replication
|
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BAC is good because it minimizes
|
the number of clones needed to make a genomic library
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Actual definition of genomic library is
|
plasmid containing clones of particular segment from the gene
|
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Plasmid can carry DNA no longer than length of
|
10 kb pairs
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BAC's are bad because they are
|
harder to work with, cut up to "subcloned" section
|
|
|
nucleic acid hybridization is when
|
you detect gene's DNA by using complementary molecule
|
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Artificial Eukaryotic chromosomes have 3 things
|
centromere, telomeres, origin of replication
|
|
|
cycle of PCR production
|
1. Reaction mixture is heated to separate DNA strands
2. Colled to allow annealing of DNA primers complementary to sequences 3.heat stable DNA polymerase extends primers in 5-3 direction |
|
|
DNA polymerase used in PCR is called
|
taq polymerase
|
|
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What is a nucleic acid probe?
|
Complementary molecule used in nucleic acid hybridization
|
|
|
Denature means
|
separate
|
|
|
Taq polymerase comes from species
|
thermus aquaticus
|
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What is an expression vector?
|
cloning vector that contains a highly active bacterial promoter sequences upstream of a restriction site where Eu. gene can be inserted
|
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Annealing means
|
hydrogen bonding
|
|
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What does RFLP stand for?
|
restriction fragment length polymorphism
|
|
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What is an RFLP?
|
when enzymes chop the genome at a certain point
|
|
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What is an SNP and what does it stand for?
|
when someone has DNA change that prevents RFLP; single nucleotide polymorphism
|
|
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What is a core sequence?
|
similar piece of DNA in satellites
|
|
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Pores in a bacteria are called
|
adhesion zones
|
|
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What parts of DNA are most susceptible to change?
|
tandem DNA
|
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DNA profiling focuses on what?
|
few variable minisatellites
|
|
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Competency must be done to _____ bacteria, ones that can go through rapid _______
|
new; growth
|
|
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What is a minisatellite?
|
tandem DNA
|
|
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What amplifies base pairs and is more effective at DNA profiling?
|
PCR
|
|
|
Lowering the temperature does what to the phosphate membrane?
|
stabilizing the negatively charged phosphate membrane
|
|
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Rapid heat addition to bacteria does what?
|
creates a temporary imbalance and current that takes DNA in
|
|
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PCR uses ____satellites
|
micro
|
|
|
What do the ions do in a salt bath for bacteria?
|
Bind to cell membrane to make it neutral
|
|
|
How does a RE help bacteria against bacteriophages?
|
cuts up foreign DNA
|
|
|
PCR stands for
|
polymerase chain reaction
|
|
|
Who invented PCR?
|
Kary Mullis
|
|
|
Restriction site is usually _____ base pairs long
|
4-8
|
|
|
microsatellites are known as
|
simple tandem repeats
|
|
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PSeudomonas are a good genus for what?
|
cleaning up chemicals
|
|
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PCR can amplify _____ base pairs
|
1000-2000
|
|
|
Microsatellites can be used to ____ & _____ the genome
|
map and sequence
|
|
|
Who wanted to originally splice cancer genes?
|
Berg
|
|
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Microsatellites are ____ than mini but just as _____
|
shorter, variable
|
|
|
antibiotics are
|
chemicals used to kill bacteria
|
|
|
cDNA is made by
|
reverse transcriptase
|
|
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With X-galactose and lacZ gene, what color signifies what?
|
no transformation= blue
transformation= white |
|
|
Nucleic Acid hybridization is
|
last step in transformation
|
|
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Who was the first genetic engineer?
|
Herb Boyer
|
|
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Transformation _____ more than it ____
|
fails, succeeds
|
|
|
cDNA is also known as
|
complementary DNA
|
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What did Boyer do? With whom?
|
put toad gene into bacteria with stan cohen
|
|
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_____ began fixing tomato crop problems
|
Horsch
|
|
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Who opposed Berg?
|
Pollack
|
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______ is the most protective form of lab wear
|
P4
|
|
|
Who was in Genetech and found insulin patch of DNA?
|
Goeddel
|
|
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Who showed recombinant DNA in system was harmless?
|
Brenner
|
|
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Who created Genetech and why?
|
Boyer and Swanson to make naturally occuring cells
|
|
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Who showed new traits could be engineered into plants?
|
Horsch
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|
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Who wanted to commercialize Boyer's work?
|
Swanson
|
|
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Who was against Genentech in insulin battle?
|
Gilbert
|
|
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How did Horsch prove work?
|
used agro bacterium to introduce new genes into plant cell
|
|
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____ are often clustered by the telomeres
|
minisatellites
|
|
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What company rules agriculture?
|
Monsanto
|
|
|
What is single nucleotide polymorphism?
|
small DNA change that stops RFLP at that site (prevents enzymes from cutting at that site)
|
|
|
PCR does what?
|
amplify DNA
|
|
|
Who invented DNA fingerprinting?
|
Professor Sir Alec Jeffreys
|
|
|
Tandem repeat DNA is what?
|
short sequence of DNA repeated many times in a row
|
|
|
______ have a repeat unit length of 6-100 bases
|
minisatellites
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What is restriction fragment length polymorphism?
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RFLP, when DNA enzymes chop short DNA sequences into pieces
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WHat is PCR?
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polymerase chain reaction
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There are 2 to several hundred repeats at a
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minisatellite
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Genetic Engineering is
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direct manipulation of genes for practical purposes
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____ have a repeat unit length of 1-7 bases
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microsatellites
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PCR works better with
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microsatellites
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_______ led to techniques needed for analyzing genes and their expressions
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recombinant DNA
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5 to 100 repeats at each _____
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microsatellite
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recombinant DNA
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DNA molecules formed when segments of DNA from two different sources are combined in vitro
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Biotechnology is
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manipulation of organimsms or components to make useful products
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_____ are scattered randomly throughout the genome
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Microsatellites
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In Vitro menas
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in test tube
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Biotechnology encompasses
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genetic engineering
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Each restriction enzyme is
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specific
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______ DNA is from another source
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foreign
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Bacteria with recombinant DNA molecule is
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recombinant bacterium
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Typical genes take up _____ of DNA in humans
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one millionth
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DNA cloning is
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methods for preparing well defined segments of DNA into multiple identical copies
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Production of multiple copies of a single gene is
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gene cloning
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restriction enzymes are also
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restriction endonucleases
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Recombinant DNA molecule is
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a plasmid that has been genetically altered with new DNA
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Gene cloning is useful because
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1. makes copies of a particular gene
2.produce a protein product |
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What are restriction endonucleases?
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enzymes that cut DNA molecules at a limited number of specific locations
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Restriction site is
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a particular short DNA sequence that is identified by specific restriction enzymes
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What is biotechnology?
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using living organisms to solve problems or create useful products
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Restriction sites are
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symmetrical
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_______ of crops have received herbicide resistant gene
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40%
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____ make recombinant DNA permanent
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DNA ligase
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Restriction fragments are
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sets of DNA that have been cut
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Sticky end is
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single stranded end left by restriction enzymes
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