lab 5

Question 1

what areas did PCR have an impact on

Answer 1

• gene mapping
• cloning
• DNA sequencing
• gene detection

Question 2

PCR template

Answer 2

any form of ds DNA

Question 3

PCR ingredients

Answer 3

• reaction buffer
• DNA subunits
• DNA polymerase
• 2 DNA primers
• template strand

Question 4

what 2 basic processes does PCR make use of

Answer 4

1. complementary DNA strand hybridization
2. DNA strand synthesis via DNA polymerase

Question 5

DNA polymerase used

Answer 5

Taq polymerase from Thermus aquaticus

Question 6

steps of PCR

Answer 6

1. denaturation
2. primer annealing
3. primer extension

Question 7

term for PCR machine

Answer 7

thermal cycler

Question 8

temperature/thermal cycling

Answer 8

rapid heating and cooling of thermal block in thermal cycler

Question 9

temp at denaturation step

Answer 9

94 degrees Celsius

Question 10

temp at annealing step

Answer 10

60 degrees Celsius

Question 11

temp at extension step

Answer 11

72 degrees Celsius

Question 12

temperature cycle =

Answer 12

denaturation + annealing + extension

Question 13

PCR generates DNA of a…

Answer 13

precise length and sequence

Question 14

which cycle are fragments of precise length generated

Answer 14

third

Question 15

what are targets of random insertions?

Answer 15

introns

Question 16

what does SINE stand for

Answer 16

short repetitive interspersed elements

Question 17

ex. of SINE

Answer 17

Alu element

Question 18

where does Alu name come from

Answer 18

Alu I restriction enzyme recognition site found in this sequence

Question 19

where are we looking for Alu sequence

Answer 19

PV92 locus on chromosome 16

Question 20

dimorphic

Answer 20

Alu element present in some ind. but not others

Question 21

homozygous Alu presence

Answer 21

941 bp

Question 22

homozygous Alu absence

Answer 22

641 bp

Question 23

heterozygous Alu

Answer 23

both 641 and 941 bp

Question 24

bioinformatics

Answer 24

discipline that integrates mathematical, statistical, and computer tools to collect/process biological data

Question 25

InstaGene matrix

Answer 25

consists of suspension of negatively charged beads which bind divalent cations - need to remove divalent cations because they will assist enzymes in DNA degradation

Question 26

supernatant

Answer 26

contains clean, intact genomic DNA

Question 27

functions of incubation steps

Answer 27

1. breakup of connective tissue 2. inactivates DNases

Question 28

master mix

Answer 28

mixture of nucleotides, buffer, and Taq DNA pol

Question 29

dye that makes DNA primers yellow

Answer 29

tartrazine

Question 30

is manual PCR possible?

Answer 30

yes