Team:Hawaii/Notebook/2008-05-30

From 2008.igem.org

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(Preparation for making competant cells)
 
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'''Grace & Norman'''
'''Grace & Norman'''
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Plated ''E. coli'' strain DH5-alpha on LB plates made yesterday
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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'''
-
 
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*Autoclaved eppendorf tubes, 10ul, 200ul, 1000uL tips for experiments during coming weeks.
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*Made [https://2008.igem.org/SOB SOB media] for growing ''E. coli'' DH5-alpha
+
*Made [[SOB|SOB media]] for growing ''E. coli'' DH5-&alpha;
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*Made SOC media
+
*Made [[SOC|SOC media]]
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*Made 20mM glucose 12.5X stock solution (12.5X 20mM = 0.25M)
+
*Made x2 bottles of 12.5X 20mM glucose stock solution (12.5X 20mM = 0.25M)
*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 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 MgSO4 for mixing post-autoclave-separately into SOB and SOC media (forgot to autoclave)
*Made stock of 1 M KOAc
*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

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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!"


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