Refer to the picture of the pGEX-6P-2 MCS sequence diagram while you answer this
ID: 3165241 • Letter: R
Question
Refer to the picture of the pGEX-6P-2 MCS sequence diagram while you answer this question. Design and write out the sequence of a pair of primers that could be used to amplify all of the DNA between the valine codon and the stop codon. Amplification of some DNA outside these regions is acceptable as part of your result. Annotate the 5’ and 3’ ends of the primers in your answer. To get good marks, the primers should adhere to the rules of good primer design. Please explain your design process as part of your answer. Include a calculation of Tm for each primer.
BamHI The multiple cloning site of pGEX-6P-2 ggt ggc gac cat cct cca aaa tog gat ctg gaa gtt ctg tte cag gg9 ccc ctg gga tccExplanation / Answer
Forward primer sequence 5' cca ccg gta gga ggt 3' . length =15 mer GC = 66.6% Tm = 50oC
Reverse primer sequence 5' tca ccg aaa cgc gcg agg 3'. length=18mer GC=66.6% Tm= 56%
Tm calculation formula= 4°C x (number of G's and C's in the primer) + 2°C x (number of A's and T's in the primer)
Justifcation for primer designing: primers are at least 15 bp apart from the site to be covered i.e valine and stop codon. GC content is in the range of 55-65 which gives optimal annealing temprature to primers. Length of primers are not less than 15 mer.
Related Questions
drjack9650@gmail.com
Navigate
Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.