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UTDallas/20 October 2010
From 2010.igem.org
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===October 20, 2010 === | ===October 20, 2010 === | ||
| - | ; | + | ;Nitrate Sensor |
*Transformed the ligated PyeaR parts | *Transformed the ligated PyeaR parts | ||
*Miniprepped, digested, ran a gel electrophoresis (took only the bottom band (~3000bp), gel purified, dephosphorylated, and then PCR purified GFP | *Miniprepped, digested, ran a gel electrophoresis (took only the bottom band (~3000bp), gel purified, dephosphorylated, and then PCR purified GFP | ||
*Ligated GFP to PalkB (the alkane promoter) | *Ligated GFP to PalkB (the alkane promoter) | ||
*Incubated PyeaR+RFP overnight | *Incubated PyeaR+RFP overnight | ||
| - | ; | + | ;Alkane Sensor |
* Midi prep part E0240 (vector) was cut with the restriction enzyme XbaI and incubated at 37C for 1 Hour | * Midi prep part E0240 (vector) was cut with the restriction enzyme XbaI and incubated at 37C for 1 Hour | ||
* Heat kill the enzyme after digestion (65C 10min) | * Heat kill the enzyme after digestion (65C 10min) | ||
Latest revision as of 20:12, 27 October 2010
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October 20, 2010
- Nitrate Sensor
- Transformed the ligated PyeaR parts
- Miniprepped, digested, ran a gel electrophoresis (took only the bottom band (~3000bp), gel purified, dephosphorylated, and then PCR purified GFP
- Ligated GFP to PalkB (the alkane promoter)
- Incubated PyeaR+RFP overnight
- Alkane Sensor
- Midi prep part E0240 (vector) was cut with the restriction enzyme XbaI and incubated at 37C for 1 Hour
- Heat kill the enzyme after digestion (65C 10min)
- Dephosphorylation of the digested vector using CIP
- Neucloetide removal after Dephosphorylation.
- Already had phosphorylated insert i.e the Palk Promoter.
- Set up overnight ligation at 12C.
