DNA recombinant II Flashcards Preview

genetics > DNA recombinant II > Flashcards

Flashcards in DNA recombinant II Deck (31):
1

if we are interested in amplifying a piece of RNA what do we use

reverse transcriptase to convert RNA into cDNA

2

When PCR continues normally we incorporate a dye what color is it and when does it start to glow

SYBR green dye, and it glows as more and more DNA is synthesized

3

DNA sequencing takes advantage of ddNTP'S what are they?

dideoxynucleotides

4

When we need to stop a DNA copy what do we use

ddNTP

5

how does a ddNTP work?

it inserts a deoxyribonucleotide with just a hydrogen bond and we need OH to polymerase

6

What do you need for DNA sequencing

1. primer
2. template (what you want to sequence)
3. Normal dNTP'S
4. Enzyme and Buffer
and now a little ddNTP'S to stop DNA extension

7

What is the future what is Pyrosequencing?

that is when the two phosphate groups detach from the triphosphate group and react chemically with enzymes giving off a light energy that shows up on a pyrosequencing machine signifying that a bond has been made and dna has been replicated

8

In pyrosequencing what does each peak mean double height, single height and none

double height is when you have two or more of the same nucleotide. ex. ccc
single height is when only one nucleotide is added.
when there are none is because it did not match up with the template strand

9

what is the quality or state of being able to assume different forms?

polymorphism such as frogs

10

when one of two or more alternative forms of an allele from a chromosomal locus resulting from differences in DNA sequences or numbers of tandem repeats-not necessarily a gene. it can be anywhere

DNA Ploymorphism

11

This is a known DNA sequence, often of DNA polymorphisms-best ones are unique in the genome, STS

DNA marker, sequence-tagged sites

12

What are the classes of DNA polymorphisms

SNP'S (single-nucleotide polymorphisms)
STR'S (short tandem repeats)
VNTR'S (variable number of tandem repeats)

13

What is southern blot?

analysis determines arrangement and location of restriction sites.

14

There are 3 blots what are their names and what are they used for?

southern blot-DNA
northern blot-DNA
western blot-proteins

15

how does southern blotting work?

dna is digested by enzymes producing fragments.
fragments are separated by size with gel electrophoresis.
dna fragments transferrred to a memebrane filter they are arranged exeactly as they are on the gel.
a labeled probe (radioactive or chemiluminescence) binds to DNA of interest.

16

This guy was a big player in the human genome project

J. Craig Venter

17

how many SNP'S were in J. Craig Venters diploid genome?

3.2 million

18

generally each gene is a STS- unique site in the genome

sequence tagged site

19

what is the most common DNA polymorphism caused by

a single point mutation of SNP'S

20

SNP'S can be harmful if they occur where? there are two places

coding genes, and regulatory regions

21

what does RFLP stand for?

Restriction Fragment Length Polymorphisms

22

what two things can be used to detect RLP'S

Southern blot or PCR

23

when were VNTR'S used to exonerate someone in a murder trial

1988

24

what was the name of the murder trials that exonerated someone using VNTR'S

Narborough murders

25

what are the positives for genetic testing

1. identifies mutations in key genes responsible for disease.
2. prenatal diagnosis
3. newborn screening-testing for genetic disease
4. carrier detection: testing of adults to calculate probability of baby having diseases-uses blood

26

why genetic testing doesn't always work

1. the gene for the disease hasnt been found.
2. the gene has many mutations making single test unreliable.
3. the presence of the gene does not always result in disease.
4. many diseases are polygenic, or caused by multiple genes

27

What takes advantage of DNA polymorphisms to identify individuals for forensics ?

DNA fingerprinting

28

most paternity tests use what to determine who the parents are

PCR

29

Gene therapy is mostly for what kind of cells

somatic

30

the procedure : take out cells, introduce normal gene (transgene) with virus vectors into them (now transgenic cell), replace in body is known as

gene therapy

31

if gene therapy is successful patients often need what?

repeated treatments