Suppose you're going to do 9 PCR reactions. The starting and final concentrations of the ingredients are listed below. Each reaction should have a volume of 50 μl. Component, stock concentration 5X Reaction Buffer MgCl2, 25 mM Nucleotide Mix, 20mM Primer 1, 10 μM Primer 2, 10 μM Taq DNA Polymerase, 5 Units/μl Template DNA, concentration unknown Final Concentration or amount 1.0X 1.5 mM 0.2 mM 1.0 μΜ 1.0 ΜΜ 1.0 Units use 1 μl per reaction (a) Briefly explain how you would set up the reactions and show all your calculations. Indicate the amounts that you would pipet into your tubes. Be clear. (Show ALL your calculations on a piece of paper, take a picture and paste it in the space below). (b) Using the amounts listed above, how many pmol of each primer will there be in each reaction? (Show ALL your calculations on a piece of paper, take a picture and paste it in the space below).

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Suppose you're going to do 9 PCR reactions. The starting and final concentrations of the
ingredients are listed below. Each reaction should have a volume of 50 μl.
Component, stock concentration
5X Reaction Buffer
MgCl2, 25 mM
Nucleotide Mix, 20mM
Primer 1, 10 μM
Primer 2, 10 μM
Taq DNA Polymerase, 5 Units/μl
Template DNA, concentration unknown
Final Concentration or amount
1.0X
1.5 mM
0.2 mM
1.0 μΜ
1.0 ΜΜ
1.0 Units
use 1 μl per reaction
(a) Briefly explain how you would set up the reactions and show all your calculations.
Indicate the amounts that you would pipet into your tubes. Be clear. (Show ALL your
calculations on a piece of paper, take a picture and paste it in the space below).
(b) Using the amounts listed above, how many pmol of each primer will there be in each
reaction? (Show ALL your calculations on a piece of paper, take a picture and paste it
in the space below).
Transcribed Image Text:Suppose you're going to do 9 PCR reactions. The starting and final concentrations of the ingredients are listed below. Each reaction should have a volume of 50 μl. Component, stock concentration 5X Reaction Buffer MgCl2, 25 mM Nucleotide Mix, 20mM Primer 1, 10 μM Primer 2, 10 μM Taq DNA Polymerase, 5 Units/μl Template DNA, concentration unknown Final Concentration or amount 1.0X 1.5 mM 0.2 mM 1.0 μΜ 1.0 ΜΜ 1.0 Units use 1 μl per reaction (a) Briefly explain how you would set up the reactions and show all your calculations. Indicate the amounts that you would pipet into your tubes. Be clear. (Show ALL your calculations on a piece of paper, take a picture and paste it in the space below). (b) Using the amounts listed above, how many pmol of each primer will there be in each reaction? (Show ALL your calculations on a piece of paper, take a picture and paste it in the space below).
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