Genetics in medicine

Question 1

recognize and cut DNA in a specific and reproducible fashion

Answer 1

class II restriction endonucleases

Question 2

restriction enzyme cut out some DNA, the top strand reads 5’ AATT 3’, what does the bottom strand read

Answer 2

3’ TTAA 5’ (cuts out palindromic sequences)

Question 3

class II restriction endonucleases leave “sticky” ends how many bps long?

Answer 3

4-8 ( 6 is most common)

Question 4

homodimers that recognize short, symmetric DNA sequences

Answer 4

restriction endonucleases

Question 5

used in Nick translation, probe prep, DNA repair & making blunt end from a sticky one

Answer 5

DNA polymerase I

Question 6

used in sanger sequencing and cloning. uses single stranded template

Answer 6

klenow fragment DNA polymerase

Question 7

the DNA pol I used in the lab is obtained from where

Answer 7

E. coli or T4 phage

Question 8

used to make DNA from RNA

Answer 8

reverse transcriptase

Question 9

DNA polymerase used in PCR

Answer 9

Taq polymerase

Question 10

type of bond ligase forms to reconnect DNA

Answer 10

phosphodiester

Question 11

single, large circular chromosomes

Answer 11

prokaryotic plamids

Question 12

3 must have features for cloning vectors

Answer 12

self replication, multiple cloning site, selectable marker

Question 13

in vitro method for rapid production of large amounts of specific DNA

Answer 13

PCR

Question 14

required for PCR

Answer 14

primer ( 20bp with free 3’ OH), heat, DNA pol I or Taq, dNTPs, MgCl, DNA and buffer

Question 15

PCR steps in order

Answer 15

1 denaturation, 2 annealing, 3 primer extension. repeat

Question 16

primer that binds to the sense strand in PCR

Answer 16

reverse. 3 to 5. (forward primer binds to antisense strand)

Question 17

polymerase used to amplify mRNA using PCR

Answer 17

reverse transcriptase

Question 18

explain the different blot types

Answer 18

SNoW DRoP

Question 19

SDS PAGE is used in which type of blot

Answer 19

western

Question 20

which blot uses a chemiluminescent substrate

Answer 20

western

Question 21

method to detect single nucleotide change

Answer 21

allele specific PCR

Question 22

test for unknown mutation

Answer 22

sequencing