User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/10/16: Difference between revisions
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# Heat for 10 minutes at 72°C. | # Heat for 10 minutes at 72°C. | ||
# Chill at 0°C until the samples needed again. | # Chill at 0°C until the samples needed again. | ||
* a new stock of luminol solution was made at a pH of 10.56. For full details on preparation, calculation, and concentration please see [[User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/10/16| Mary's Notebook]] | |||
==Conclusions== | |||
* The goal for today was the run the HRP chemiluminescence assay. Last week there had been no emittance from the sample. The luminol pH had been at 8 and after researching the literature, we decided that it was necessary to increase the pH of luminol in order to push forward a reaction. As a result, we aimed to make a luminol solution with a pH of 10.5. The actual pH of the luminol was 10.56. | |||
* We also decided that we needed to increase the concentration of the HRP in the cuvet. The Fluorimeter was not working and so we just used visual analysis to see if any fluorescence was occurring. The first concentration in the cuvet we tested was 2mM- which did not produce any luminescence. The next concentration we attempted was 10mM which also did not result in any luminescence. We then decided to double this final concentration to 20mM HRP while maintaining 3mM concentration of Luminol. This sample resulted in fluorescence of the luminol in the form of a blue light! We plan on running different variations of the HRP concentration beginning with 20mM tomorrow when conducting the chemiluminescence assay. | |||
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Revision as of 18:10, 23 October 2012
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PCR of ADA MutationsGoals
Preparation
Concentration of plasmid needed for experiment= 100ng/μL
.56mg= 5600 ng M1V1= M2V2 5600 ng/μL x V1= 100ng x 1000μL
V2= 17.857μL is the amount of plasmid solution pipetted out in order to get a concentration of 100ng/μL Procedure
Conclusions
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