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40 Cards in this Set

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Molecular cloning allows for the creation of two types of libraries. the 2 types of libraries are ... and ...
genomic DNA library
cDNA library
this DNA library represents expressed genes only:
cDNA
the construct the genomic library, genomic DNA is partially digested into segments. the segments must represent ... and each segment must ... another genomic fragment in the library
the entire genome
overlap
in constructing a cDNA library, the source is total ... from a specific tissue or developmental stage of embryogenesis
RNA
in creating a cDNA library, after extraction of total RNA, the next step is to ...
Isolate the polyadenylated mRNA
in constructing a cDNA library, after isolating the polyadenylated mRNA, you will need to use ... to convert the mRNA into ds cDNA
reverse transcriptase
in constructing a cDNA library, after you have you double-stranded cDNA, you will want to do what with it?
Clone the cDNAs into vectors
what is so special about a vector that contains a Shine Delgarno sequence?
it is an expression vector that will express the protein being cloned
In a cDNA library, the genes transcribed at a ... frequency will be represented more often than those transcribed at a lower frequency.
higher
... are short sequences of single stranded DNA that are complimentary to DNA sequences of specific alleles and can be radioactively labeled and used as probes to detect single nucleotide changes.
Allele-specific oligonucleotides (ASO)
Any one of a series of two or more different genes that occupy the same position (locus) on a chromosome.
allele
what is the advantage of Allele-specific oligonucleotides (ASO) analysis over Southern blot analysis?
ability to detect single nucleotide changes
RFLP stands for ...
Restriction Fragment Length Polymorphism
the following describes what:
Genetic variations at the site where a restriction enzyme cuts a piece of DNA. Such variations affect the size of the resulting fragments. These sequences can be used as markers on physical maps and linkage maps.
RFLP
A common variation in the sequence of DNA among individuals.
polymorphism
there are 2 types of RFLP's. briefly describe each:
1. mutation leads to creation or deletion of restriction site

2. Alteration of genetic material leads to change in size of restriction site
what is an indel?
a mutation that inserts or deletes nucleotides into/from a sequence
what is a point mutation?
a mutation that replaces one of the nucleotides
the following describes what:
the synthesis of new DNA strands on a single stranded template and the random incorporation of chain-terminating nucleotide analogues.
Sanger method
what is needed for DNA sequencing:
1.
2.
3.
4.
5.
1. ssDNA template
2. labeled primer
3. DNA polymerase
4. the four dNTPs
5. 4 separate reaction tubes containing one of four
dideoxynucleotides (ddNTPs) - chain terminators
in DNA sequencing, what type of gel must be used?
denaturing polyacrylamide gel
... exploits highly variable repeating sequences and based on the idea that an individual will have a distinct pattern of these variable repeating sequences
DNA fingerprinting
VNTR stands for:
variable number of tandem repeats
minisatellite
repeat is 9-65 nucleotide in length
microsatellite
di-, tri- or tetranucleotide repeats
what are the basic steps for DNA fingerprinting?
total genomic DNA digested and Southern blotted. use probe against the repetitive sequence (if there is limited amount of DNA available, PCR)
briefly describe the microarray technique:
isolate mRNA from cell
use this mRNA as templates to generate cDNA with a "fluorescent tag"
labeled samples are then mixed and incubated with a microarray containing the immobilized genes A, B, C, and D
labeled molecules bind to the sites on the array corresponding to the genes expressed in each cell.
why would one use the microarray technique?
analyze expression of many genes in a single experiment
if you test healthy tissue vs diseased tissue using microarray, what might you tell about the diseased tissue?
which genes are turned ON/OFF in response to disease
gene expression pattern of a disease state
microarrays can be used for comparative genomic hybridization which is used to detect ...
genomic loss or gain
change in the number of copies of a particular gene involved in a disease
comparative genomic hybridization can be used to monitor mutations related to cancer progression
false
mutation microarray analysis can look at single nucleotide ... associated with particular disease
polymorphisms
introduction of a new gene into an organism refers to ...
transgenic
the following describes what:
Genes responsible for particular traits or disease susceptibility are chosen and extracted. Next they are injected into fertilized mouse eggs. Embryos are implanted in the uterus of a surrogate mother. The selected genes will be expressed by some of the offspring.
transgenic mice
what is a knockout mouse?
genetically engineered mouse that has had one or more of its genes made inoperable
what is the purpose of a knockout mouse?
determine possible function of normal gene in whole animal
... is the random insertion of mutant gene in genome and is useful when mutant gene ... function of normal gene
gene addition
overrides
transgenic mouse-TDD expresses defective version of ... impacting both transcription and DNA repair and mimics mutation in humans that causes ...
XPD gene (TFIIH)
trichothiodystrophy (TDD)
transgenic mouse-TDD suffers from ...
premature aging
what is the correlation between transgenic mouse-TDD what happens in humans?
accumulation of DNA damage contributes to aging process