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Team:Brown/Notebook/July19
From 2010.igem.org
Monday, July 19 2010
Results of yesterday’s plates:
Amp+ (older batch of plates): BBa_B0015 Terter: very sparse growth – about 7 colonies. No contamination.
Amp+ (older batch of plates): BBa_R0080 pAraC: about 50 to 70 colonies, no contamination.
Amp+ (new batch of plates): BBa_1732100 LacI: About 30 colonies, but with contamination aroud the edges of the plate – looks like a lawn of bacteria and there is a little contamination in the middle of the plate.
Kan+ BBa_C0080 AraC: LAWN. Looks like there may be something strange going on with our Kan plates…
Note here to make more Kan plates. Also, the new batch of Amp+ plates appears to have side/edge of the plate contamination.
Plates
- Kan+ -- Bba_K19105 -- Lovtap reporter 2 -- Lawn. Defective kan plates? looks like null
- null -- Bba_K19105 -- Lovtap reporter 2 -- Lawn. Identical to kan plate
- tet+ 10ug/ml -- Bba_K19105 -- Lovtap reporter 2 -- Beginnings of a lawn
- amp+ -- Bba_K19105 -- Lovtap reporter 2 -- ~100 colonies, successful :)
- amp+ -- TrpR promoter -- Many colonies (~1000)
- kan+ -- TrpR promoter -- lawn
- null -- TrpR promoter -- lawn
- tet+ 10ug/ml -- TrpR promoter -- semi-lawn
Harvesting plates
11AM:
- Picked colonies (random 2 from each plate
- Put into 15mL culture tube
- 3ml of lb+3ul amp stock at 100mg/mL)
- Incubate at 37C
- Did not bother plating/picking a colony from kan+ plate, there were no colonies to pick
New Part
Rehydrated Bba_J04421 (LacI reporter - CFP construct) from 2008 registry
- Made 5ul of DNA from 1 paper punch
- Added all 5ul to 150ul of XL1
- Plated on amp+ plate
Transform: Ligation of WillRS+pGEM
XL1Blues
- Followed modified Adrian protocol
- Added 110mL LB
- Added 50ul XGAL from Adrian
- Added 2ul IPTG
- Put on new amp plaets
- Incubated overnight at 37C
Note - The new Amp plates seen to be slightly softer than normal. This may be due to how they were stored after cooling.
