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Lab 10 entry- PCR Reactions

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
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Lab 9 entry

Once my DNA was extracted, I prepped it for gel electrophoresis.

  1. First, we dotted ~2mL of blue tracking dye on a piece of parafilm.
  2. Then, I put ~3mL of each extracted DNA on top of a dot, changing tips every time.
  3. Finally, I put my pipette on 6mL and moved the solution from the parafilm into the wells of the gel.
  4. We ran the gel.