Lab 7

Question 1

transformation efficiency,

Answer 1

which is a measure of the success of the transformation process

Question 2

2. Know how to calculate and explain transformation efficiency

Answer 2

Colonies/microgram/dilution

Question 3

o Do you know the components of PCR

Answer 3

Template DNA, forward and reverse primers, DNTPs, Taq polymerase, MgCl2, buffer

Question 4

Polymerase Chain Reaction (PCR),

Answer 4

which is a method to amplify millions of copies of DNA from a very small amount of starting material.

Question 5

denaturation

Answer 5

break the hydrogen bonds between complementary bases, resulting in two single-stranded DNA templates that can be copied.

Question 6

annealing,

Answer 6

lowers the temperature to between 30-65°C to allow primers to bind to complementary regions on the DNA template

Question 7

elongation, or extension,

Answer 7

where deoxyribonucleoside triphosphates (dATP, dGTP, dCTP, dTTP; collectively ‘dNTPs’) are added to the growing DNA chain in a complementary and antiparallel manner by a DNA polymerase enzyme at 72 degrees c

Question 8

o Do you know how to perform PCR?

Answer 8

Denaturation, annealing, extension in thermocycler for 30 cycles in MgCl2 buffer

Question 9

o Do you understand what a primer is?

Answer 9

Short, single-stranded segments of DNA that bind to beginning and end of sequence to be amplified