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

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(Continuation of Chemiluminescence)
(Continuation of Chemiluminescence)
Line 31: Line 31:
* The molarity of sodium carbonate (MW 105.9784 g/mol) added was calculated:  
* The molarity of sodium carbonate (MW 105.9784 g/mol) added was calculated:  
<math>\frac{1.91 g}{15 mL}</math> × <math>\frac{1 mol}{105.9784 g}</math> = <math>\frac{0.00120 mol}{mL}</math>
<math>\frac{1.91 g}{15 mL}</math> × <math>\frac{1 mol}{105.9784 g}</math> = <math>\frac{0.00120 mol}{mL}</math> × <math>\frac{1 mL}{1E-3 L}</math> = <math>\frac{1.20 mol}{L}</math>

Revision as of 02:33, 26 October 2012

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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

  • The luminol prepared from the previous chemiluminescence laboratory period had a pH of 7 to 8. Reviewing a journal article of Xiaoyu, it was determined that the optimal pH of luminol was 12.5. As a result, it was decided to prepare a new solution of luminol at a pH of 10 to 11.
  • A weight of .0112 g of luminol was added to 6 mL of water. The buffer composed of a direct addition of .0737 g of sodium carbonate and .4358 g of sodium bicarbonate.
  • Using a pH meter, the electrode detected the pH at 8.73. As suggested by Dr. Hartings, a solution of sodium carbonate was made to increase the pH of the solution.
  • Several adjustments were made in increasing the pH. A total weight of 1.91 g of sodium carbonate dissolved in 15 mL of water was added to the 6 mL solution of luminol.
  • The molarity of sodium carbonate (MW 105.9784 g/mol) added was calculated:

\frac{1.91 g}{15 mL} × \frac{1 mol}{105.9784 g} = \frac{0.00120 mol}{mL} × \frac{1 mL}{1E-3 L} = \frac{1.20 mol}{L}

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