Haynes Lab:Notebook/Engineering PC-TFs/2015/02/23: Difference between revisions
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| Total || 25.0 μL || 25μL | | Total || 25.0 μL || 25μL | ||
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| colspan="3" | Mix the reaction(s) thoroughly by flicking the tube.<br> Placed in thermocycler on LCR setting. | | colspan="3" | Mix the reaction(s) thoroughly by flicking the tube.<br> Placed in thermocycler on LCR setting. *Volume reduced to ~10μL after removal from PCR machine. Settings incorrect? | ||
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*DH5α-T cells were thawed in ice. | *DH5α-T cells were thawed in ice. | ||
*1.5mL tubes were setup and labeled. | *1.5mL tubes were setup and labeled. | ||
*60μL of thawed cells were mixed and then transferred into each | *60μL of thawed cells were mixed and then transferred into each tube. | ||
*The ligation products were then transferred into their respective tubes and flicked to mix. Set back on ice. | *The ligation products were then transferred into their respective tubes and flicked to mix. Set back on ice. | ||
*All tubes incubated on ice for 35 minutes. | *All tubes incubated on ice for 35 minutes. | ||
*All tubes were heat shocked on the heat block at 42°C for 30 seconds and then placed back on ice for 2 minutes. | *All tubes were heat shocked on the heat block at 42°C for 30 seconds and then placed back on ice for 2 minutes. | ||
*900μL SOC Medium was pipetted into each | *900μL SOC Medium was pipetted into each tube. | ||
*Tubes were taped into an empty plate. The plate was taped to the shaker within the incubator. | *Tubes were taped into an empty plate. The plate was taped to the shaker within the incubator. | ||
*Incubated on shaker for 1 hour at 37°C and 240rpm. | *Incubated on shaker for 1 hour at 37°C and 240rpm. |
Latest revision as of 00:47, 27 September 2017
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SummaryLCR LCR Calculations
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