The primers for amplification of the ALDH promoter sequences have arrived. These primers have the biobrick overhangs. Primers were diluted and the first PCR reaction to amplify the ALDH promoters was set up and run overnight. Primer stocks were diluted to 100mM and then working concentrations of 10µM were made up. 100µM stocks were produced using the IDT re-suspension calculator. PCR was set up using Pfu polymerase. Template used was whole cells (mammalian). Cells were thawed, spun down and the supernatant removed and then re-suspended in 50uL DMSO. PCR was left to run overnight. Annealing temperature used was 54 degrees, this is slightly higher than the annealing temperature generally used in the lab. poster Retrieved from ""