PANGILINAN, Kylela P.

MODX311 (LAB)
BSMD 2Y3-5(B)

UNIT TASK #3: Pipetting exercise, Reagent Preparation, Use of pH meter

Critical Thinking Questions:

1. You are tasked to prepare a Master Mix using the BGI PCR Kit. You have 25
samples to process. Since you are only using a half-plate you only need 3 controls. You
are only a beginner in reagent preparation and therefore aren't that much confident on
your pipetting skills, so you created an additional number for pipetting error which is 2.

BGI Kit Master Mix:

Reaction Mix per well: 18.5μL
Enzyme Mix per well: 1.5μL
Master Mix per well: 20μL

CALCULATE FOR THE:

A. Desired volume for reaction mix (5 points)
Total number of samples: 25
Total number of controls: 3
Total number for pipetting error: 2
Total: 30

A: 18.5uL x 30 = 555μL
 
B. Desired volume for enzyme mix (5 points)
A: 1.5uL x 30 = 45μL

C. Master mix volume (5 points)

A: 555uL + 45uL = 600μL
2. Discuss the process of your reagent preparation on Question 1. (10 points)

A: First step in preparing the reagent is to calculate the amount of reaction and
enzyme mix needed based on the number of samples, controls and pipetting errors that
you have prepared. After successfully figuring out the needed volume of the mixes,
aspirate the reaction mix (555μL), and dispense into a microcentrifuge tube. Same goes
with the enzyme mix. Draw the needed volume (45μL) and dispense into the same
microcentrifuge that was used. Then, use a vortex mixer to incorporate the solutions.
You now have your master mix.