A sample that was not in the strip of cDNA tubes was not qPCRd in the last run (20110518). Ran that sample (labelled 2GE R 1/5), as well as positive control DNA (07-CB-719-3), 1GE (cDNA) and 3GE (cDNA). Master mix calcs are here. Plate layout, cycling params, etc. can be found in the qPCR Report (see Results).
Tested Colleen’s newly designed OsHV primers, which were based off of SOLiD next-gen sequencing expression data. Three primer sets (OsHV_ORF87, OsHV_ORF104, and OsHV_ORF107) were tested on two different DNA samples recommended by Colleen (07-CB-719-1 & 07-CB-719-3; both from 10/19/07). Master mix was set up according to Colleen’s protocol. Master mix can be found here. Plate layout, cycling params, etc. can be found in the qPCR Report (see Results).
Results show good amplification and clean melt curves with primers OsHV_ORF87 and 107. OsHV_ORF104 shows good amplification, but multiple peaks in the melt curve. Will run qPCR on cDNAs using primers OsHV_ORF87 and 107. Will discuss with Colleen and Carolyn whether they want to try optimizing the reaction for primer set OsHV_ORF104.