Team:Hawaii/Notebook/2008-05-30
From 2008.igem.org
(Difference between revisions)
(→Preparation for making competant cells) |
(→Preparation for making competant cells) |
||
(4 intermediate revisions not shown) | |||
Line 4: | Line 4: | ||
'''Grace & Norman''' | '''Grace & Norman''' | ||
- | Plated ''E. coli'' strain DH5-alpha on LB plates made yesterday | + | Plated ''E. coli'' strain DH5-α on LB plates made yesterday |
:*one plate was plated using the hockey puck method for maximum growth | :*one plate was plated using the hockey puck method for maximum growth | ||
:*one plate was plated using the 4-way streak method to isolate colonies | :*one plate was plated using the 4-way streak method to isolate colonies | ||
- | Plates were placed in a 37C incubator for 5 hours then transfered to a 4C incubator for storage. | + | Plates were placed in a 37C incubator for <strike>5 hours then transfered to a 4C incubator for storage.</strike> 12 hours then transferred to 26C incubator for growth till Sunday 3pm. (colonies were too small) |
== Preparation for making competant cells == | == Preparation for making competant cells == | ||
''' Grace, Margaret, & Norman''' | ''' Grace, Margaret, & Norman''' | ||
- | + | *Autoclaved eppendorf tubes, 10ul, 200ul, 1000uL tips for experiments during coming weeks. | |
- | *Made [ | + | *Made [[SOB|SOB media]] for growing ''E. coli'' DH5-α |
- | *Made SOC media | + | *Made [[SOC|SOC media]] |
- | *Made 20mM glucose solution | + | *Made x2 bottles of 12.5X 20mM glucose stock solution (12.5X 20mM = 0.25M) |
- | *Made CMB80 media | + | *Made CMB80 media <strike>(need to filter in detergent free bottle tomorrow)</strike> (already filtered into plastic sterile bottle that came with the filtering kit, stored @ 4C) |
- | *Made stock of 1 M | + | *Made MgSO4 for mixing post-autoclave-separately into SOB and SOC media (forgot to autoclave) |
+ | *Made stock of 1 M KOAc | ||
*Made stock of 0.1 N HCl | *Made stock of 0.1 N HCl | ||
Latest revision as of 04:40, 3 June 2008
Projects | Events | Resources | ||
---|---|---|---|---|
Sponsors | Experiments | Milestones | Protocols | |
Notebook (t) | Meetings (t) |
Contents |
Plated E. coli
Grace & Norman
Plated E. coli strain DH5-α on LB plates made yesterday
- one plate was plated using the hockey puck method for maximum growth
- one plate was plated using the 4-way streak method to isolate colonies
Plates were placed in a 37C incubator for 5 hours then transfered to a 4C incubator for storage. 12 hours then transferred to 26C incubator for growth till Sunday 3pm. (colonies were too small)
Preparation for making competant cells
Grace, Margaret, & Norman
- Autoclaved eppendorf tubes, 10ul, 200ul, 1000uL tips for experiments during coming weeks.
- Made SOB media for growing E. coli DH5-α
- Made SOC media
- Made x2 bottles of 12.5X 20mM glucose stock solution (12.5X 20mM = 0.25M)
- Made CMB80 media
(need to filter in detergent free bottle tomorrow)(already filtered into plastic sterile bottle that came with the filtering kit, stored @ 4C) - Made MgSO4 for mixing post-autoclave-separately into SOB and SOC media (forgot to autoclave)
- Made stock of 1 M KOAc
- Made stock of 0.1 N HCl
Discussion
Grace, Margaret, and Norman will come in on Sunday at 3pm to grow up E. coli into exponential phase in preparation for inducing chemical competency.
Quote of the Day
- Dr. Presting: "Wow, I didn't know three people could fit in this room."
- M: "Well, not maybe three of you."
- Dr. Presting: "Are you calling me fat?"
- M: "No, no. You're tall, dark, and handsome!"
[http://manoa.hawaii.edu/ ][http://manoa.hawaii.edu/ovcrge/ ][http://www.ctahr.hawaii.edu ]