QUOTE (postdoc @ Mar 29 2005, 02:27 PM)
I am interested to know how you prepare your PCR stock solution to minimizing pipetting, do you put two primers in a single stock solution? Do you put all except primers in a stock solution?
It mostly depends on the question asked / purpose of the PCR. If we just want to check the presence of a gene or if PCR is possible without using the PCR product for further downstream processes then we use already pre-made PCR master mixes or PCR beads where you just have to add your primers and template (and water if using beads). If I am using the PCR product for sequencing than I am using a proof-reading enzyme....in that case I am mixing the PCR solution with the individual ingredients when I need it. However, the primer sets we keep separate and only add them to the stock when they are needed, especially because we are working with several different primer sets (and sometimes mix and match different primers).