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From 2010.igem.org

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Revision as of 16:41, 23 September 2010

Contents 1 Monday 1.1 69. Expt: Growing up of cell cultures of above experimental results & of results on p49 (Will) 1.2 70. Expt: Miniprep pSB1C3 from colonies (Anja) 1.3 71. Expt: Gibson assembly of pBAD, luxCD,AB,EG and pSB1C3 (Anja)

Monday

Result (from Expt. 68):

No growth overnight, 2 growths out of 3 plates (+1 fungus) after weekend of growing. No glow.

69. Expt: Growing up of cell cultures of above experimental results & of results on p49 (Will)

Strains G1, j1 and G28wh were grown up in 5ml LB + 2µl Chloramphenicol, and on plates containing LB+Cm.

Left to grow at 30°C for 24h.

Results: Strains grew but did not glow after 48h.

70. Expt: Miniprep pSB1C3 from colonies (Anja)

Suspended a 'streak' of bacteria transformed with BBa_J04450 (registry part in pSB1C3) in 250µl buffer P1. Followed 'Bench protocol: Qiagen Spin Miniprep Kit Using a Microcentrifuge'.

Nanodrop 12.8ng/µl

71. Expt: Gibson assembly of pBAD, luxCD,AB,EG and pSB1C3 (Anja)

PCR:

Template	Primer f.	Primer r.	Amplified Fragment
I0500	prefix.f.pBadstart	luxCstart.r.pBADend	pBAD (A)
pJS555	pBADend.f.luxCstart	luxAstart.r.luxDend	luxCD (B)
pJS555	luxDend.f.luxAstart	luxEstart.r.luxBend	luxAB (C)
pJS555	luxBend.f.luxEstart	suffix.rev.luxGend	luzEG (D)
BBa_J04450	luxGend.for.suffix	pBADstart.r.prefix	pSB1C3 (E)

BBa_J04450: once purified (E), once colony PCR (EC)