Haynes Lab:Notebook/Engineering PC-TFs/2014/05/22: Difference between revisions
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==Summary== | ==Summary== | ||
'''Long Transformation'''<br> | '''Long Transformation'''<br> | ||
Using the set up ligation products from 5/21. | Using the set up ligation products from 5/21. | ||
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*50μL of thawed cells were transferred into each of these three empty tubes and pipetted to mix. | *50μL of thawed cells were transferred into each of these three empty tubes and pipetted to mix. | ||
* The two ligation setup products were then transferred into tubes one and two respectively and flicked to mix. Set back on ice. | * The two ligation setup products were then transferred into tubes one and two respectively and flicked to mix. Set back on ice. | ||
* A 1:1000 RhlI plasmid was used as a positive control; 1μL of this plasmid was added to 19μL dH<sub>2</sub>O to equal the other two products 20μL total volume. Added to the third tube with 50μL thawed cells. | * A 1:1000 RhlI plasmid was used as a positive control; 1μL of this plasmid was added to 19μL dH<sub>2</sub>O to equal the other two products' 20μL total volume. Added to the third tube with 50μL thawed cells. | ||
*All tubes incubated on ice for 30 minutes. | *All tubes incubated on ice for 30 minutes. | ||
*All three tubes were heat shocked on the heat block at 42°C for 30 seconds and then placed back on ice for 2 minutes. | *All three tubes were heat shocked on the heat block at 42°C for 30 seconds and then placed back on ice for 2 minutes. |
Revision as of 16:02, 22 May 2014
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SummaryLong Transformation
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