Mary set up a similar PCR reaction. The Tm of the forward and reverse
primers she uses is 55oC. The conditions that were optimized by her
instructor were as follows:
Cycle Temperature (oC) Time
1X 94o 3 minutes
30X 94o 30 seconds Denaturation
50o 30 seconds Annealing
72o 60 seconds Extension
Mary sets up the PCR, but decides to see if she can get better results
than her instructor, and makes the following changes in separate PCR
runs.
Describe the effect that each of her changes would have on the amount
and purity of her PCR product. (Briefly explain).
She changes the annealing temperature to 60oC
She increases the number of cycles to 31
She changes the magnesium concentration from 1.0 mM to 2.0 mM
She adds non-complementary restriction enzyme sequences to the 3’ ends
of the primers
Biology Homework Solutions
#152529
PCR
Mary set up a similar PCR reaction. The Tm of the forward and reverse primers she uses is 55oC. The conditions that were optimized by her instructor were as follows:
Cycle
Temperature (oC)
Time
1X
94o
3 minutes
30X
94o
30 seconds
Denaturation
50o
30 seconds
Annealing
72o
60 seconds
Extension
Mary sets up the PCR, but decides to see if she can get better results than her instructor, and makes the following changes in separate PCR runs.
Describe the effect that each of her changes would have on the amount and purity of her PCR product. (Briefly explain).
A.She changes the annealing temperature to 60oC
B.She increases the number of cycles to 31
C.She changes the magnesium concentration from 1.0 mM to 2.0 mM
D.She adds non-complementary restriction enzyme sequences to the 3’ ends of the primers
Answers to several questions regarding PCR.
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