Illinois/1 August 2008

Receptor Tyrosine Kinase Group

PCR troubleshooting of yesterday's reaction: use gradient for annealing temp (40-65 degrees), use more template, leave out end primers

Four series of reactions using a gradient annealing temp were run in addition to a fifth series omitting the end primers.

annealing temps: 40, 43.8, 50, 54, 60, and 64 degrees

PCR program: 1. 94 degrees 5 min 2. 94 degrees 1 min 3. annealing step 1 min 4. 72 degrees 1 min 5. GOTO 2. 29 cycles 6. HOLD at 4 degrees

PCR products run on 1.5% agarose gel at 200V for ~20 minutes, then 240V for ~20 minutes.

Products in the 700-800bp range in all lanes. Perhaps it worked because of using a differnt product from the 7-30 PCR.