User:Tamra L. Fisher/Notebook/Research for Dr. Hartings/2012/08/15: Difference between revisions
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==A Note About Last Night's Ligation== | ==A Note About Last Night's Ligation== | ||
There seems to have been a power outage last night so the thermocycler was not on and not at 16°C when I came in this morning. The total ligation time was 15 hours and 55 minutes, but I am not sure how much of that time was spent at 16°C and how much of that was spent at room temperature. I have realized that I don't have enough double-digested | There seems to have been a power outage last night so the thermocycler was not on and not at 16°C when I came in this morning. The total ligation time was 15 hours and 55 minutes, but I am not sure how much of that time was spent at 16°C and how much of that was spent at room temperature. I have realized that I don't have enough double-digested pQE-80 to re-do the ligation tonight, but I will begin digesting the plasmid and still amplify and transform last night's product. | ||
==Amplification of Last Night's Ligation Product== | ==Amplification of Last Night's Ligation Product== |
Revision as of 12:56, 15 August 2012
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A Note About Last Night's LigationThere seems to have been a power outage last night so the thermocycler was not on and not at 16°C when I came in this morning. The total ligation time was 15 hours and 55 minutes, but I am not sure how much of that time was spent at 16°C and how much of that was spent at room temperature. I have realized that I don't have enough double-digested pQE-80 to re-do the ligation tonight, but I will begin digesting the plasmid and still amplify and transform last night's product. Amplification of Last Night's Ligation ProductThis procedure was done for the wild-type Asc Hb that was ligated with pQE-80-L-Kan last night. The pQE-80 primer was used:
Most components for this PCR came from the Phusion® High-Fidelity PCR Kit. Cutting with HindIII for CloningThis was done with the pQE-80-L-Kan midiprep from 06/19/12.
Running an Analytical DNA Gel
[[Image: Transformations
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