Edinburgh/12 August 2008

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Edinburgh iGEM 2008

June
M	T	W	T	F	S	S
						[http://2008.igem.org/wiki/index.php?title=Edinburgh/1_June_2008&action=edit 1]
[http://2008.igem.org/wiki/index.php?title=Edinburgh/2_June_2008&action=edit 2]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/3_June_2008&action=edit 3]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/4_June_2008&action=edit 4]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/5_June_2008&action=edit 5]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/6_June_2008&action=edit 6]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/7_June_2008&action=edit 7]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/8_June_2008&action=edit 8]
[http://2008.igem.org/wiki/index.php?title=Edinburgh/9_June_2008&action=edit 9]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/10_June_2008&action=edit 10]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/11_June_2008&action=edit 11]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/12_June_2008&action=edit 12]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/13_June_2008&action=edit 13]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/14_June_2008&action=edit 14]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/15_June_2008&action=edit 15]
[http://2008.igem.org/wiki/index.php?title=Edinburgh/16_June_2008&action=edit 16]	[http://2008.igem.org/Edinburgh/17_June_2008 17]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/18_June_2008&action=edit 18]	[http://2008.igem.org/Edinburgh/19_June_2008 19]	[http://2008.igem.org/Edinburgh/20_June_2008 20]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/21_June_2008&action=edit 21]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/22_June_2008&action=edit 22]
[http://2008.igem.org/wiki/index.php?title=Edinburgh/23_June_2008&action=edit 23]	[http://2008.igem.org/Edinburgh/24_June_2008 24]	[http://2008.igem.org/Edinburgh/25_June_2008 25]	[http://2008.igem.org/Edinburgh/26_June_2008 26]	[http://2008.igem.org/Edinburgh/27_June_2008 27]	[http://2008.igem.org/Edinburgh/28_June_2008 28]	[http://2008.igem.org/Edinburgh/29_June_2008 29]
[http://2008.igem.org/Edinburgh/30_June_2008 30]

July
M	T	W	T	F	S	S
	[http://2008.igem.org/Edinburgh/1_July_2008 1]	[http://2008.igem.org/Edinburgh/2_July_2008 2]	[http://2008.igem.org/Edinburgh/3_July_2008 3]	[http://2008.igem.org/Edinburgh/4_July_2008 4]	[http://2008.igem.org/Edinburgh/5_July_2008 5]	[http://2008.igem.org/Edinburgh/6_July_2008 6]
[http://2008.igem.org/Edinburgh/7_July_2008 7]	[http://2008.igem.org/Edinburgh/8_July_2008 8]	[http://2008.igem.org/Edinburgh/9_July_2008 9]	[http://2008.igem.org/Edinburgh/10_July_2008 10]	[http://2008.igem.org/Edinburgh/11_July_2008 11]	[http://2008.igem.org/Edinburgh/12_July_2008 12]	[http://2008.igem.org/Edinburgh/13_July_2008 13]
[http://2008.igem.org/Edinburgh/14_July_2008 14]	[http://2008.igem.org/Edinburgh/15_July_2008 15]	[http://2008.igem.org/Edinburgh/16_July_2008 16]	[http://2008.igem.org/Edinburgh/17_July_2008 17]	[http://2008.igem.org/Edinburgh/18_July_2008 18]	[http://2008.igem.org/Edinburgh/19_July_2008 19]	[http://2008.igem.org/Edinburgh/20_July_2008 20]
[http://2008.igem.org/Edinburgh/21_July_2008 21]	[http://2008.igem.org/Edinburgh/22_July_2008 22]	[http://2008.igem.org/Edinburgh/23_July_2008 23]	[http://2008.igem.org/Edinburgh/24_July_2008 24]	[http://2008.igem.org/Edinburgh/25_July_2008 25]	[http://2008.igem.org/Edinburgh/26_July_2008 26]	[http://2008.igem.org/Edinburgh/27_July_2008 27]
[http://2008.igem.org/Edinburgh/28_July_2008 28]	[http://2008.igem.org/Edinburgh/29_July_2008 29]	[http://2008.igem.org/Edinburgh/30_July_2008 30]	[http://2008.igem.org/Edinburgh/31_July_2008 31]

August
M	T	W	T	F	S	S
				[http://2008.igem.org/Edinburgh/1_August_2008 1]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/2_August_2008&action=edit 2]	[http://2008.igem.org/Edinburgh/3_August_2008 3]
[http://2008.igem.org/Edinburgh/4_August_2008 4]	[http://2008.igem.org/Edinburgh/5_August_2008 5]	[http://2008.igem.org/Edinburgh/6_August_2008 6]	[http://2008.igem.org/Edinburgh/7_August_2008 7]	[http://2008.igem.org/Edinburgh/8_August_2008 8]	[http://2008.igem.org/Edinburgh/9_August_2008 9]	[http://2008.igem.org/Edinburgh/10_August_2008 10]
[http://2008.igem.org/Edinburgh/11_August_2008 11]	[http://2008.igem.org/Edinburgh/12_August_2008 12]	[http://2008.igem.org/Edinburgh/13_August_2008 13]	[http://2008.igem.org/Edinburgh/14_August_2008 14]	[http://2008.igem.org/Edinburgh/15_August_2008 15]	[http://2008.igem.org/Edinburgh/16_August_2008 16]	[http://2008.igem.org/Edinburgh/17_August_2008 17]
[http://2008.igem.org/Edinburgh/18_August_2008 18]	[http://2008.igem.org/Edinburgh/19_August_2008 19]	[http://2008.igem.org/Edinburgh/20_August_2008 20]	[http://2008.igem.org/Edinburgh/21_August_2008 21]	[http://2008.igem.org/Edinburgh/22_August_2008 22]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/23_August_2008&action=edit 23]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/24_August_2008&action=edit 24]
[http://2008.igem.org/Edinburgh/25_August_2008 25]	[http://2008.igem.org/Edinburgh/26_August_2008 26]	[http://2008.igem.org/Edinburgh/27_August_2008 27]	[http://2008.igem.org/Edinburgh/28_August_2008 28]	[http://2008.igem.org/Edinburgh/29_August_2008 29]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/30_August_2008&action=edit 30]	[http://2008.igem.org/wiki/index.php?title=Edinburgh/31_August_2008&action=edit 31]

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Week 9

Tuesday 12 August 08

Plate 115 made with subs from L33 (Pl. 109/110: pSB1A2+pCstA), L34 (Pl. 111/112: pSB1A2+dxs+crtE) and L35 (Pl. 113/114: pSB1A2+dxs+lims1) (CF)
Over-night cultures of L33 (pSB1A2+pCstA, sub-cloned on Plate 115), L34 (pSB1A2+dxs+crtE, sub-cloned on Plate 115) and L35 (pSB1A2+dxs+lims1, subcloned on Plate 115) set up, ready for minipreps tomorrow.
M121-M123: Minipreps of pSB1A2+crtY (1,2,3). M124-M126: Minipreps of pSB1A2+rbs+crtY (1,2,3). note: I messed up M121 by adding 200µl instead of 100µl of sol 1. They are in the green box, the orange one is full. (OG)
Double digests of M121 to M126, run on Gel 46. It looks good! M121 is good! (Yan)
Purified P61 (rrnB) for sequencing. (CF/AM)
Re-purified already purified genomic DNA from C. fimi. This meaningless endeavour resulted from a misunderstanding. Hopefully, the doubly-purified DNA will still be useable. (AM)
P57 (cex from cells) EcoRI/PstI double digest, undigested P64 (cex from genomic DNA), P66 (su1) and P67 (iso2) run on Gel 45. P57 double digest yielded no clear bands; P64 yielded a clear band at 1.5 kb. (CF/AM)
Purified P57 (cex from cell suspension) and P64 (cex from genomic DNA). Digested both with PstI. (AM)
PCR for cenA from C. fimi genomic DNA (P65): denaturing 1 min, annealing 56°C for 20s, extension 30s, with glycerol. (AM)
P57 and P64 (cex) PstI digests and P65 (cenA) run on Gel 47. Results indicate success! (AM)
Re-did the excise of crtE from BABEL2 (M79); cut out the band responsible for crtE and Edinbrick 1 on the gel, followed by putification of crtE and Edinbrick1 from the cut-out gel. Finally, ligation of crtE to Edinbrick1 (L38). (HX, AH)
Analytical digests as follows (AH):
- M109 and M110 (pSB1A2+crtB+crtI): BamHI/EcoRV
- M115 and M116 (pSB1A2+glgC-mut1,2): HindIII
- M115 and M116 (pSB1A2+glgC-mut1,2): EcoRI/HindIII
Digests of M109, M110, M115 and M116 were run on gel 48. M109 and M110 should give two bands if both genes are present. HindIII digest of M115/116 should give a single band indicating linearised plasmid with glgC. Double digest of M115/116 should give an insert band of ~1kb and a vector + ~500bp of insert band. Results do not look good. Bands seem to indicate much too large products, but will discuss with Chris tomorrow.(AH)
M116 (pSB1A2+glgC-mut1,2) and M120 (BABEL2+glgC-mut1,2,3) submitted for sequencing. (CF)

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