User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/10/16

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PCR mutation

  • In reference to the PCR Mutation protocol, 100 ng/μL of the primer was needed for the reaction. The weight of the primer in the provided container was 0.46 mg. A ratio of the weight over volume was equated to the required concentration of the primer:

0.46 mg = 0.46E6 ng

\frac{0.46E6 ng}{x \mu L} = \frac{100 ng}{1 \mu L} of primer in water = 4600 μL of water

  • There is limited space in the plastic container (1 mL). Instead of dissolving 0.46E6 ng of the primer in 4600 μL of water, the entire primer was dissolved in 1 mL water.
  • Using M1V1 = M2V2, the volume taken from the solution of 0.46E6 ng in 1 mL of water was calculated to be 217.39 μL. This was transferred to a new tube and filled up with water to a total volume of 1 mL.

V1 = \frac{100 ng/ \mu L  *  1000 \mu L}{0.46E3 ng/ \mu L} = 217.39 μL of the dissolved primer in water

  • The procedure listed in PCR Mutation protocol was strictly followed. After the addition of all reagents, the sample was placed in the thermocycler.

Continuation of Chemiluminescence

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