"
Team:Yale/Notebook/Week2
From 2011.igem.org
(→Notebook: Week 2) |
(→Notebook: Week 2) |
||
| Line 8: | Line 8: | ||
- Made TB buffer (DH5-alpha competent cell buffer). 5mLs of 1M HEPES buffer (final concentration 10mM), 75mL of 0.1M CaCL2*2H2O (final concentration 15mM), 9.32g KCl (final concentration 250mM) was added to a beaker. Volume was brought to 300mL. pH was brought to 6.7 with 6.25M KOH. 5.44g of MnCL2*4H2O (final concentration 55mM) was added, and the volume was brought to 500mL. The contents of the beaker were sterile filtered into a sterile bottle purchased from the basement store. | - Made TB buffer (DH5-alpha competent cell buffer). 5mLs of 1M HEPES buffer (final concentration 10mM), 75mL of 0.1M CaCL2*2H2O (final concentration 15mM), 9.32g KCl (final concentration 250mM) was added to a beaker. Volume was brought to 300mL. pH was brought to 6.7 with 6.25M KOH. 5.44g of MnCL2*4H2O (final concentration 55mM) was added, and the volume was brought to 500mL. The contents of the beaker were sterile filtered into a sterile bottle purchased from the basement store. | ||
| - | - Made 500mL LB buffer, which was distributed in 2x250mL Erlenmeyer flasks and 2x500mL Erlenmeyer flasks. No antibiotic was added. | + | - Made 500mL LB buffer (no agar), which was distributed in 2x250mL Erlenmeyer flasks and 2x500mL Erlenmeyer flasks. No antibiotic was added. |
| - | + | - Added 250ul of frozen DH5-alpha cells to 50ml culture flask. Incubated on shaker at 37 overnight. | |
| - | + | ''Tuesday'' | |
| - | + | - Transferred 4mL of overnight culture to 200mL tube. Waited 5 hours before checking the OD. The OD was >2 but it should be 0.55. Will innoculate another 50mL culture overnight but wait shorter amount of time. | |
Revision as of 17:25, 24 May 2011
Notebook: Week 2
Monday
- Meeting scheduled in the Med School TAC building with Martin and Girish tomorrow at 2:30pm about using their plasmids with the anti-freeze glycoprotein that they isolated. - Meeting scheduled for Friday with Farren at 2:30pm in West Campus, building B-31, Room 209.
- Made TB buffer (DH5-alpha competent cell buffer). 5mLs of 1M HEPES buffer (final concentration 10mM), 75mL of 0.1M CaCL2*2H2O (final concentration 15mM), 9.32g KCl (final concentration 250mM) was added to a beaker. Volume was brought to 300mL. pH was brought to 6.7 with 6.25M KOH. 5.44g of MnCL2*4H2O (final concentration 55mM) was added, and the volume was brought to 500mL. The contents of the beaker were sterile filtered into a sterile bottle purchased from the basement store.
- Made 500mL LB buffer (no agar), which was distributed in 2x250mL Erlenmeyer flasks and 2x500mL Erlenmeyer flasks. No antibiotic was added.
- Added 250ul of frozen DH5-alpha cells to 50ml culture flask. Incubated on shaker at 37 overnight.
Tuesday
- Transferred 4mL of overnight culture to 200mL tube. Waited 5 hours before checking the OD. The OD was >2 but it should be 0.55. Will innoculate another 50mL culture overnight but wait shorter amount of time.
Wednesday
Plan:
Accomplished:
Thursday
Plan:
Accomplished:
Friday
Plan:
Accomplished:
