When primers (oligonucleotides) arrive at the lab from thecompany, they are typically shipped dry, and we need to resuspendthem in an aqueous solution before use. We usually add a volume ofTE buffer or water for resuspension to obtain a final concentrationof 100 μM (stock solution). As you can see in thisprotocol, the working solution of each primeris 2 μM (see the table below).

 

Indicate your calculations to prepare a workingsolution of the reverse primer at a concentration of 2 μM from thestock solution, consider the volume of primer needed per PCRreaction (per student), and that there are 24 students in your labsection

Reagent: 5x PCR buffer (Containing 15 mM MgCl2) deoxyribonucleotide triphosphate (dNTPs) (10 mm) Forward primer (2 M) pMCT118-f Reverse primer (2 M) pMCT118-r ultrapure H20 (nuclease-free) Taq DNA polymerase (5 U/L).-GoTaq G2 DNA pol TOTAL Volume 10 L 1 L 5 L 5 L 23.8 L 0.2 45 L MasterMix (per bench: 4 students: x 4.2) 42 L 4.2 21 L 21 99.96 0.84 L 189