You want to amplify the DNA between the stretch of sequence shown in the figure
ID: 13518 • Letter: Y
Question
You want to amplify the DNA between the stretch of sequence shown in the figure below. Of the listed primers chose the pair that will allow you to amplify the DNA by PCR.
A. Forward - 5’-ATGGACTAGGGCCTAATCGGT-3’
Reverse - 5’-CATAGGTACGTAATATACTACTT-3’
B. Forward - 5’-ATGGACTAGGGCCTAATCGGT-3’
Reverse - 5’-TTCATCATATAATGCATGGATAC-3’
C. Forward - 5’- ACCGATTAGGCCCTAGTCCAT-3’
Reverse - 5’-AAGTAGTATATTACGTACCTATG-3’
D. Forward - 5’-ATGGACTAGGGCCTAATCGGT-3’
Reverse - 5’-AAGTAGTATATTACGTACCTATG-3’
Explanation / Answer
The answer is C. You need to choose primers that are complimentary to the known sequences IN THE CORRECT ORIENTATION. This means the 5' end of your primer needs to line up with the 3' end of your known DNA. If we look at C in more detail: First, ignore the answer choices. You need to write out your own corresponding sequences first. Forward Primer Selection: Known DNA: 5'-ATGGACTAGGGCCTAATCGGT-3' Primer DNA: 3'-TACCTGATCCCGGATTAGCCA-5' If you flip the primer DNA sequence around to read 5' to 3', you get the answer in choice C. Reverse Primer Selection: Known DNA: 5'-CATAGGTACGTAATATACTACTT-3' Primer DNA: 3'-GTATCCATGCATTATATGATGAA-5' Again, if you flip the primer DNA sequence around to read 5' to 3', you get the answer in choice C. C is the answer! Hope this helps.
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