This week we ran the PCR reactions for our Mimulus guttatus project – materials shown below. We made enough Mastermix for 18 reactions even though we only had 12- just to make sure we had enough.
| Ingredients | Per Reaction | [18 reactions] |
| ddH2O | 13.36 | 240.48 |
| 10x buffer | 2.00 | 36 |
| MgCl2 | 2.00 | 36 |
| BSA | 1.00 | 18 |
| dNTPs | 0.20 | 3.6 |
| F-primer | 0.20 | 3.6 |
| R-primer | 0.20 | 3.6 |
| Taq | 0.04 | .72 àRounded to .80 (for pipette) |
| Template | 1.00 | n/a |
| Total | 20.00 |
We labeled our tubes as shown below:
| Tubes | Specimen ID |
| QS 1 MM | MM1 |
| QS 2 MM | MM2 |
| QS 3 MM | MM3 |
| QS 4 OY | OY01 |
| QS 5 OY | OY02 |
| QS 6 OY | OY03 |
| QS 7 | EB |
| QS 8 | RDRK001 |
| QS 9 | SHOR005 |
| QS 10 | KRS1 |
| QS 11 | KRS2 |
| QS 12 | KRS3 |
We added 19 microliter of Mastermix to each of the 12 reactions, mixed them on the vortex then Prof. Paul ran the PCR reaction for us.
