Molecular & Cellular Biology Lab Instructor: Elmar Schmid, Ph.D. (Student Name) Primer Design Worksheet A B 5’ – T G C G T T A G G C G C G A A T T G T C A T G G T T A C G C C G T A A G G C T A T A G – 3’ C D 3’ – A C G C A A T C C G C G C T T A A C A G T A C C A A T G C G G C A T T C C G A T A T C – 5’ 1. 2. 3. 4. 5. Write the nucleotide sequence of one suitable forward and reverse primer pair into the empty white boxes below the selected target sequences A, B C or D (grey boxes) Indicate the 5’ and 3’ ends of your selected primer pair Draw arrows to indicate the polymerization direction of the Taq polymerase after annealing of your chosen primer pair to the template DNA What would be the length (in bp) of the PCR amplified DNA using your selected pair above? ________ bp What would be the calculated melting temperatures Tm of the two chosen (forward and reverse) primers? (see “PCR” lab introduction section on my website for help) Tm (forward) = ___ oC 6. Tm (reverse) = ___ oC Which annealing temperature TA would you chose to do your PCR with the two primers above? (See “PCR” lab introduction section on my website for help) TA = ___ oC