The Polymerase Chain Reaction, or PCR, is a technique used to make copies of DNA. Specifically, PCR uses an enzyme known as a polymerase, which adds individual DNA bases to a growing chain of DNA. The PCR reaction is dependent upon the primers, short strands of DNA the polymerase needs to begin reproducing DNA, and the primer matrix, a mixture of reagents and the primers themselves. Each PCR reaction should be a total volume of no more than 50 uL.
- 10x PCR Buffer
- 2mM dNTP
- 50pmol primers
- 25mM MgCl2
- 1 pmol template DNA
- Purified water
- Eppendorf tubes
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Instructions
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1
Add 2.5 uL of the PCR buffer to an Eppendorf tube, as well as 2.5 uL of the dNTPs.
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2
Add 2 uL (50 pmol) of the primers to the Eppendorf tube and 1 uL of MgCl2.
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3
Add water to reach a final volume of 50 uL. Add 1 pmol of the template DNA, or 1 uL of DNA solution, whichever is easier to measure. Your primer matrix is prepared.
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4
Repeat Steps 1 through 3 for as many matrices as you want to prepare.
