BASIL2022GV3HDT: Difference between revisions

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===='''Coupled kinase assay'''====

Two rounds of coupled kinase assays were run using 3HDT with ATP and dCMP as substrates. The concentration of dCMP was 109mM. The first round of assay (3 total assays) used 5μL of 3HDT and various amounts of 109mM of dCMP. 6.88μL of dCMP resulted in the highest specific activity (0.37669 U/mg) and increasing the substrate amount ~2μL had a similar but slightly less specific activity of 0.3268 U/mg. However, when we repeated the first kinase assays we did (using 5μL of 3HDT), the results ~~did not turn out the same or even similar to the first round~~ indicating a potential error ~~could have happened~~. In addition, the protein in the second round was older (original protein but about a week old from when it was made) which could have affected the specific activity with the substrate because the protein was starting to expire/decrease function.

Two rounds of coupled kinase assays were run using 3HDT with ATP and dCMP as substrates. The concentration of dCMP was 109mM. The first round of assay (3 total assays) used 5μL of 3HDT and various amounts of 109mM of dCMP. 6.88μL of dCMP resulted in the highest specific activity (0.37669 U/mg) and increasing the substrate amount ~2μL had a similar but slightly less specific activity of 0.3268 U/mg. However, when we repeated the first kinase assays we did (using 5μL of 3HDT), there are discrepancies in the results indicating a potential experimental error such as not pipetting up and down to mix, bubbles, taking too long between mixing the substrate in and reading the plate. In addition, the protein in the second round was older (original protein but about a week old from when it was made) which could have affected the specific activity with the substrate because the protein was starting to expire/decrease function.

BASIL2022GV3HDT: Difference between revisions

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Revision as of 03:28, 24 April 2022 view source Autumn Forrester (talk \| contribs) 241 edits No edit summary ← Older edit		Revision as of 04:16, 24 April 2022 view source Autumn Forrester (talk \| contribs) 241 edits No edit summary Newer edit →
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	===='''Coupled kinase assay'''====		===='''Coupled kinase assay'''====
	Two rounds of coupled kinase assays were run using 3HDT with ATP and dCMP as substrates. The concentration of dCMP was 109mM. The first round of assay (3 total assays) used 5μL of 3HDT and various amounts of 109mM of dCMP. 6.88μL of dCMP resulted in the highest specific activity (0.37669 U/mg) and increasing the substrate amount ~2μL had a similar but slightly less specific activity of 0.3268 U/mg. However, when we repeated the first kinase assays we did (using 5μL of 3HDT), the results ~~did not turn out the same or even similar to the first round~~ indicating a potential error ~~could have happened~~. In addition, the protein in the second round was older (original protein but about a week old from when it was made) which could have affected the specific activity with the substrate because the protein was starting to expire/decrease function.		Two rounds of coupled kinase assays were run using 3HDT with ATP and dCMP as substrates. The concentration of dCMP was 109mM. The first round of assay (3 total assays) used 5μL of 3HDT and various amounts of 109mM of dCMP. 6.88μL of dCMP resulted in the highest specific activity (0.37669 U/mg) and increasing the substrate amount ~2μL had a similar but slightly less specific activity of 0.3268 U/mg. However, when we repeated the first kinase assays we did (using 5μL of 3HDT), there are discrepancies in the results indicating a potential experimental error such as not pipetting up and down to mix, bubbles, taking too long between mixing the substrate in and reading the plate. In addition, the protein in the second round was older (original protein but about a week old from when it was made) which could have affected the specific activity with the substrate because the protein was starting to expire/decrease function.