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295 Cards in this Set

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