Ran PCRs for the following samples:
| Date | Product | Gene | Starting Plasmid | Forward Primer | Reverse Primer | Thermocycler Variables: # cycles / annealing temp / extension time | Link to Gel Picture |
|---|---|---|---|---|---|---|---|
| 6/23 | Q2-BACE2-QX | BACE2 | pEXPR BACE2 | iGEM 016 BACE2 primer forward | iGEM 017 BACE2 primer reverse | (60)30 / 60C / 30s | Picture 6/23 |
| 6/23 | Q1-eYFP-Q2 | eYFP | pEXPR TRE:eYFP | iGEM 018 eYFP primer forward | iGEM 019 eYFP primer reverse | (60)30 / 60C / 30s | Picture 6/23 |
| 6/23 | Q1-BACE2-Q2 | BACE2 | pEXPR BACE2 | iGEM 001-2 BACE2_Primer_F | iGEM 002-2 BACE2_Primer_R | (60)30 / 60C / 30s | Picture 6/23 |
| 6/23 | Q2-eYFP-QX | eYFP | pEXPR TRE:eYFP | iGEM 003-2 YFP_Primer_F | iGEM 004-2 YFP_Primer_R | (60)30 / 60C / 30s | Picture 6/23 |
Ran PCRs according to protocol on NEB website: https://www.neb.com/protocols/2012/09/06/protocol-phusion-high-fidelity-pcr-master-mix-with-hf-buffer-m0531
...
| STEP | TEMP | TIME |
| Initial Denaturation | 98°C | 30 seconds |
| 30 Cycles | 98°C 60°C 72°C | 5 seconds 30 seconds 30 seconds |
| Final Extension | 72°C | 5 minutes |
| Hold | 4°C |