We used our DNA template to run PCR reactions (20mL). To do this, we created a master mix with the following solutions and measurements:
Ingredients | per rxn | per 18rxns (mL) |
ddH20 | 13.36 | 240.48 |
10x buffer | 2.00 | 36.0 |
MgCl2 | 2.00 | 36.0 |
BSA | 1.00 | 18.0 |
dNTPs | 0.20 | 3.60 |
F-primer | 0.20 | 3.60 |
R-primer | 0.20 | 3.60 |
Taq | 0.04 | 0.80 |
Template | 1.00 | n/a |
Total | 20.00 | 19.00/rxn |
First, I labeled three tubes with tube names QS10-12 respectively. Then, I used a pipette to put 19mL of the master mix in each tube. I then put each DNA template into its respective tube, changing pipette tips. Below is a list of tube names and their respective specimen ID.
Tubes | Specimen ID |
QS1MM | MM1 |
QS2MM | MM2 |
QS3MM | MM3 |
QS4OY | OY01 |
QS5OY | OY02 |
QS6OY | OY03 |
QS7 | EB |
QS8 | RDRK001 |
QS9 | SHOR005 |
QS10 | KRS1 |
QS11 | KRS2 |
QS12 | KRS3 |