DNA recombinant II

Question 1

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

Answer 1

reverse transcriptase to convert RNA into cDNA

Question 2

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

Answer 2

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

Question 3

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

Answer 3

dideoxynucleotides

Question 4

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

Answer 4

ddNTP

Question 5

how does a ddNTP work?

Answer 5

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

Question 6

What do you need for DNA sequencing

Answer 6

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

Question 7

What is the future what is Pyrosequencing?

Answer 7

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

Question 8

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

Answer 8

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

Question 9

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

Answer 9

polymorphism such as frogs

Question 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

Answer 10

DNA Ploymorphism

Question 11

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

Answer 11

DNA marker, sequence-tagged sites

Question 12

What are the classes of DNA polymorphisms

Answer 12

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

Question 13

What is southern blot?

Answer 13

analysis determines arrangement and location of restriction sites.

Question 14

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

Answer 14

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

Question 15

how does southern blotting work?

Answer 15

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.

Question 16

This guy was a big player in the human genome project

Answer 16

J. Craig Venter

Question 17

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

Answer 17

3.2 million

Question 18

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

Answer 18

sequence tagged site

Question 19

what is the most common DNA polymorphism caused by

Answer 19

a single point mutation of SNP’S

Question 20

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

Answer 20

coding genes, and regulatory regions

Question 21

what does RFLP stand for?

Answer 21

Restriction Fragment Length Polymorphisms

Question 22

what two things can be used to detect RLP’S

Answer 22

Southern blot or PCR

Question 23

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

Answer 23

1988

Question 24

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

Answer 24

Narborough murders

Question 25

what are the positives for genetic testing

Answer 25

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

Question 26

why genetic testing doesn’t always work

Answer 26

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

Question 27

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

Answer 27

DNA fingerprinting

Question 28

most paternity tests use what to determine who the parents are

Answer 28

PCR

Question 29

Gene therapy is mostly for what kind of cells

Answer 29

somatic

Question 30

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

Answer 30

gene therapy

Question 31

if gene therapy is successful patients often need what?

Answer 31

repeated treatments