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Team:Cornell/Notebook/Salicylate reporter
From 2012.igem.org
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(→June 29th, Friday) |
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* Gel purified PCR product. | * Gel purified PCR product. | ||
** Quantified product at 22.4 ng/uL | ** Quantified product at 22.4 ng/uL | ||
| - | ** Set up digestion of PCR product with EcoRI and AscI. | + | ** Set up digestion of PCR product with EcoRI-HF and AscI. |
| + | *** 22 ng/uL --> 45.5 uL sample for 1 ug digest | ||
| + | *** Used 5.3 uL NEB Buffer 4 | ||
| + | *** 1 uL of each enzyme, diluted to 1 unit/uL | ||
* [[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 | ||
Revision as of 01:11, 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.
- Quantified product at 22.4 ng/uL
- Set up digestion of PCR product with EcoRI-HF and AscI.
- 22 ng/uL --> 45.5 uL sample for 1 ug digest
- Used 5.3 uL NEB Buffer 4
- 1 uL of each enzyme, diluted to 1 unit/uL
- 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)
