Lab

Question 1

What are the 3 steps of PCR?

Answer 1

1) denaturation (hot)
2) annealing (cool)
3) extension/synthesis (hot)

Question 2

What does the denaturation step of PCR entail?

Answer 2

splits DNA strands

Question 3

What does the annealing step of PCR entail?

Answer 3

primers bind to DNA strands

Question 4

What does the extension/synthesis step of PCR entail?

Answer 4

primers elongated

Question 5

What is the function of the Taq polymerase in PCR? Why is a bacteria like E coli not used?

Answer 5

elongates primers added during annealing by adding complementary nucleotides; thermophilicity of exremophile required

Question 6

PCR Reaction Component: Taq polymerase

Function: ?

Answer 6

adds complementary nucelotides

Question 7

PCR Reaction Component: PCR buffer

Function: ?

Answer 7

stabilize DNA and other components

Question 8

PCR Reaction Component: Primer

Function: ?

Answer 8

short DNA sequences that allow Taq to elongate DNA

Question 9

PCR Reaction Component: dNTPs

Function: ?

Answer 9

complementary nucleotides added by Taq used to elongate primers

Question 10

PCR Reaction Component: MgCl2

Function: ?

Answer 10

required for Taq polymerase

Question 11

PCR Reaction Component: ? Function: adds complementary nucelotides

Answer 11

Taq polymerase

Question 12

PCR Reaction Component: ?

Function: stabilize DNA and other components

Answer 12

PCR buffer

Question 13

PCR Reaction Component: ?

Function: short DNA sequences that allow Taq to elongate DNA

Answer 13

Primer

Question 14

PCR Reaction Component: ?

Function: required for Taq polymerase

Answer 14

MgCl2

Question 15

Formula for total copies created using PCR

Answer 15

DNA(total)=DNA(copies)x2^n

n=# of cycles

Question 16

How are DNA fragments separated using agarose gel electrophoresis?

Answer 16

DNA fragments are separated based on charge and size (small move quicker)

Question 17

What is the purpose of adding ethidium bromide to the gel?

Answer 17

binds to DNA and flueorscences udner UV light

Question 18

Why was sample loading buffer added to your samples before loading it into the well in the agarose gel?

Answer 18

To increase the desnity of DNA fragments to prevent mizing DNA with the buffer

Question 19

How can you determine the molecular weight of DNA fragments using agarose gel electrophoresis?

Answer 19

smaller fragments move faster than larger fragments

Question 20

What is taxonomy?

Answer 20

the science of namin and grouping organisms based on genetic relationships?

Question 21

How does taxonomy differ for eukaryotes vs prokaryotes?

Answer 21

eukaryotes- sexual reproduction

prokaryotes- similar characteristic

Question 22

The identification of bacteria is aided by a multivolume identification manual known as

Answer 22

Manual of Systematic Bacteriology

Question 23

What is the Manual of Systematic Bacteriology?

Answer 23

multi-volume manual use in the identification of bacteria

Question 24

Identification of bacteria typically begins with ____

Answer 24

gram stain and morphological characteristics

Question 25

A catalase test can be used to separate ____ bacteria from ____ bacteria

Answer 25

gram positive; gram negative