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Team:Cornell/Notebook/Salicylate reporter
From 2012.igem.org
(Difference between revisions)
(→June 29th, Friday) |
(→June 29th, Friday) |
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| Line 11: | Line 11: | ||
* Gel purified PCR product (DPW) | * Gel purified PCR product (DPW) | ||
** Quantified product at 22.4 ng/uL | ** Quantified product at 22.4 ng/uL | ||
| - | ** Set up digestion of PCR product with EcoRI-HF and AscI. | + | ** Set up digestion of p21 PCR product with EcoRI-HF and AscI. |
*** 22 ng/uL --> 45.5 uL sample for 1 ug digest | *** 22 ng/uL --> 45.5 uL sample for 1 ug digest | ||
*** Buffer 4 | *** Buffer 4 | ||
** Ran digestion on gel. | ** Ran digestion on gel. | ||
| + | ** Sliced out relevant band on gel, stored overnight at -20. | ||
* [[Miniprep]]ped overnight cultures of PL14-PL20 (STC) | * [[Miniprep]]ped overnight cultures of PL14-PL20 (STC) | ||
** Using C1015 rotor, 6666 x g, the Corning culture tubes only fit in the centrifuge with the lids off | ** Using C1015 rotor, 6666 x g, the Corning culture tubes only fit in the centrifuge with the lids off | ||
| Line 26: | Line 27: | ||
** Amplifying nah operon from Gibson 1 | ** Amplifying nah operon from Gibson 1 | ||
** Appending BioBrick cutsites for ligation into pSB3C5 | ** Appending BioBrick cutsites for ligation into pSB3C5 | ||
| + | |||
| + | ====June 30th, Saturday==== | ||
| + | * Took PCR out of thermal cycler at 9:00 am (DPW) | ||
| + | ** Set of gel using NEB 100bp and 2-log ladders (10:00am) | ||
| + | * Continued gel extraction of p21 PCR digest from previous day (SS) | ||
Revision as of 14:51, 30 June 2012
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June
Week
June 29th, Friday
- Vent PCR at 11:00 (DPW)
- Dylan's magic triple anneal method (55/60/63)
- Gel purified PCR product (DPW)
- Quantified product at 22.4 ng/uL
- Set up digestion of p21 PCR product with EcoRI-HF and AscI.
- 22 ng/uL --> 45.5 uL sample for 1 ug digest
- Buffer 4
- Ran digestion on gel.
- Sliced out relevant band on gel, stored overnight at -20.
- Miniprepped overnight cultures of PL14-PL20 (STC)
- Using C1015 rotor, 6666 x g, the Corning culture tubes only fit in the centrifuge with the lids off
- Made LB, 3x 60 mL in 100 mL bottles (SS)
- Made LB Agar, 4 x 250 mL LB Agar in 500 mL bottles (CS)
- Autoclaved LB, LB Agar, and milliQ (SS)
- Made LB plates with Kan (CMR)
- CUGEM movie outing at 8:00 pm.
- Phusion PCR at 10:00 pm (DPW)
- Dylan's magic triple anneal method (57/65/70, 35 cycles total)
- Amplifying nah operon from Gibson 1
- Appending BioBrick cutsites for ligation into pSB3C5
June 30th, Saturday
- Took PCR out of thermal cycler at 9:00 am (DPW)
- Set of gel using NEB 100bp and 2-log ladders (10:00am)
- Continued gel extraction of p21 PCR digest from previous day (SS)
