"
Team:Cornell/Notebook/Salicylate reporter
From 2012.igem.org
(Difference between revisions)
(→June 30th, Saturday) |
(→Week) |
||
| Line 17: | Line 17: | ||
** Ran digestion on gel. (~11:00 pm) | ** Ran digestion on gel. (~11:00 pm) | ||
** Sliced out relevant band on gel, stored overnight at -20. | ** Sliced out relevant band on gel, stored overnight at -20. | ||
| - | + | <br> | |
| - | + | ||
* [[Miniprep]]ped overnight cultures of PL14-PL20 (~1:00 pm, STC) | * [[Miniprep]]ped overnight cultures of PL14-PL20 (~1:00 pm, 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 | ||
| - | + | <br> | |
* Made [[LB]], 3x 60 mL in 100 mL bottles (~ 3:00pm, SS) | * Made [[LB]], 3x 60 mL in 100 mL bottles (~ 3:00pm, SS) | ||
* Made [[LB|LB Agar]], 4 x 250 mL LB Agar in 500 mL bottles (~3:00, CS) | * Made [[LB|LB Agar]], 4 x 250 mL LB Agar in 500 mL bottles (~3:00, CS) | ||
* [[Autoclave]]d LB, LB Agar, and milliQ (~3:30 pm, SS) | * [[Autoclave]]d LB, LB Agar, and milliQ (~3:30 pm, SS) | ||
* Made [[LB]] plates with Kan (~6:30 pm, CMR) | * Made [[LB]] plates with Kan (~6:30 pm, CMR) | ||
| - | + | <br> | |
| - | + | ||
* CUGEM movie outing at 8:00 pm. | * CUGEM movie outing at 8:00 pm. | ||
| - | + | <br> | |
| - | + | ||
* [[Phusion PCR]] at 10:00 pm (DPW) | * [[Phusion PCR]] at 10:00 pm (DPW) | ||
** Dylan's magic triple anneal method (57/65/70, 35 cycles total) | ** Dylan's magic triple anneal method (57/65/70, 35 cycles total) | ||
** 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 | ||
| - | + | <br> | |
====June 30th, Saturday==== | ====June 30th, Saturday==== | ||
* Took PCR out of thermal cycler at 9:00 am (DPW) | * Took PCR out of thermal cycler at 9:00 am (DPW) | ||
Revision as of 15:13, 30 June 2012
| Home | Team | Project | Parts | Modeling | Notebook | Protocols | Safety | Attributions |
|---|
|
Write description
June
Week
June 29th, Friday
- Vent PCR at 11:00 (DPW)
- Amplifying both previous Phusion PCR band and original p21 template
- Dylan's magic triple anneal method (55/60/63)
- Gel purified PCR product from Phusion template (~1:20 pm) (DPW)
- Quantified product at 22.4 ng/uL
- Set up digestion of p21 PCR product with EcoRI-HF and AscI (~9:00 pm).
- 22 ng/uL --> 45.5 uL sample for 1 ug digest
- Buffer 4
- Ran digestion on gel. (~11:00 pm)
- Sliced out relevant band on gel, stored overnight at -20.
- Miniprepped overnight cultures of PL14-PL20 (~1:00 pm, 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 (~ 3:00pm, SS)
- Made LB Agar, 4 x 250 mL LB Agar in 500 mL bottles (~3:00, CS)
- Autoclaved LB, LB Agar, and milliQ (~3:30 pm, SS)
- Made LB plates with Kan (~6:30 pm, 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)
- Set up ligation of p21 PCR digest extraction and dephosphorylated pBBRBB+mtrB (11:11am, SS)
