Ran qPCR on a set of previously unopened box of Immomix (lot #112E) to verify that all vials in the box are good, due to the variability we have experienced within lot numbers in our most recent troubleshooting of the withering syndrome qPCR assay.
The nine vials in the box were numbered one through nine and then individual master mixes were made from each with the SPUD internal amplification control (1:10 million dilution as template) assay.
All samples were run in triplicate.
Master mix calcs are here: 20140124 – qPCR Immomix Checks
See the qPCR Report in the Results below for plate layout, cycling params, etc.
Vial 5 is the ONLY vial that performed as expected! A single vial out of 10 vials! Vial 3 is potentially useable.
All other vials showed very little amplification, as well as variation in their amplification. Performance was absolutely dreadful. Considering these were reactions where the master mixes were simply distributed into wells, the variation seen in some of them is baffling.
Will contact Bioline about lot number expiration dates, as well as getting reimbursed (in Immomix) for all the time/money we’ve wasted troubleshooting.