Imperial College/22 September 2008

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Revision as of 17:07, 23 September 2008

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

September
M	T	W	T	F	S	S
[http://2008.igem.org/Imperial_College/1_September_2008 1]	[http://2008.igem.org/Imperial_College/2_September_2008 2]	[http://2008.igem.org/Imperial_College/3_September_2008 3]	[http://2008.igem.org/Imperial_College/4_September_2008 4]	[http://2008.igem.org/Imperial_College/5_September_2008 5]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/6_September_2008&action=edit 6]	[http://2008.igem.org/Imperial_College/7_September_2008 7]
[http://2008.igem.org/Imperial_College/8_September_2008 8]	[http://2008.igem.org/Imperial_College/9_September_2008 9]	[http://2008.igem.org/Imperial_College/10_September_2008 10]	[http://2008.igem.org/Imperial_College/11_September_2008 11]	[http://2008.igem.org/Imperial_College/12_September_2008 12]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/13_September_2008&action=edit 13]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/14_September_2008&action=edit 14]
[http://2008.igem.org/Imperial_College/15_September_2008 15]	[http://2008.igem.org/Imperial_College/16_September_2008 16]	[http://2008.igem.org/Imperial_College/17_September_2008 17]	[http://2008.igem.org/Imperial_College/18_September_2008 18]	[http://2008.igem.org/Imperial_College/19_September_2008 19]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/20_September_2008&action=edit 20]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/21_September_2008&action=edit 21]
[http://2008.igem.org/Imperial_College/22_September_2008 22]	[http://2008.igem.org/Imperial_College/23_September_2008 23]	[http://2008.igem.org/Imperial_College/24_September_2008 24]	[http://2008.igem.org/Imperial_College/25_September_2008 25]	[http://2008.igem.org/Imperial_College/26_September_2008 26]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/27_September_2008&action=edit 27]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/28_September_2008&action=edit 28]
[http://2008.igem.org/Imperial_College/29_September_2008 29]	[http://2008.igem.org/Imperial_College/30_September_2008 30]

October
M	T	W	T	F	S	S
		[http://2008.igem.org/Imperial_College/1_October_2008 1]	[http://2008.igem.org/Imperial_College/2_October_2008 2]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/3_October_2008&action=edit 3]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/4_October_2008&action=edit 4]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/5_October_2008&action=edit 5]
[http://2008.igem.org/wiki/index.php?title=Imperial_College/6_October_2008&action=edit 6]	[http://2008.igem.org/Imperial_College/7_October_2008 7]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/8_October_2008&action=edit 8]	[http://2008.igem.org/Imperial_College/9_October_2008 9]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/10_October_2008&action=edit 10]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/11_October_2008&action=edit 11]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/12_October_2008&action=edit 12]
[http://2008.igem.org/Imperial_College/13_October_2008 13]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/14_October_2008&action=edit 14]	[http://2008.igem.org/Imperial_College/15_October_2008 15]	[http://2008.igem.org/Imperial_College/16_October_2008 16]	[http://2008.igem.org/Imperial_College/17_October_2008 17]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/18_October_2008&action=edit 18]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/19_October_2008&action=edit 19]
[http://2008.igem.org/wiki/index.php?title=Imperial_College/20_October_2008&action=edit 20]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/21_October_2008&action=edit 21]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/22_October_2008&action=edit 22]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/23_October_2008&action=edit 23]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/24_October_2008&action=edit 24]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/25_October_2008&action=edit 25]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/26_October_2008&action=edit 26]
[http://2008.igem.org/wiki/index.php?title=Imperial_College/27_October_2008&action=edit 27]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/28_October_2008&action=edit 28]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/29_October_2008&action=edit 29]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/30_October_2008&action=edit 30]	[http://2008.igem.org/wiki/index.php?title=Imperial_College/31_October_2008&action=edit 31]

Wet Lab

Miniprep

9 minipreps were carried out for samples 5-1, 5-2, 8-1, 8-2, 8-3, 9-2, 9-3, 12-2 and 12-3.
Minipreps 8-1, 8-2 and 8-3 digested with EcoRI and SpeI

Lane 1 - Undigested miniprep 8-1, lane 2 - digested miniprep 8-1, lane 3 - undigested miniprep 8-2, lane 4 - digested miniprep 8-2, lane 5 - undigested miniprep 8-3 and lane 6 - digested miniprep 8-3.

The vector in mini-preps 8-1 and 8-2 appear to be ok (~3kb (pSB1AK3)), although there no insert is visible (this may be due to it being too small however). The vector and possible insert in midiprep 8-3 appear to be the wrong size.

Single Colony PCR

A variety of ligations were carried out last week and to test there success we carried out single colony PCR using Psb primers to verify if an insert is present in the vector AK3. For each ligation thress seperate colonies were picked to carry out SCP and a negative colony where no DNA was present. The gel below shows the resuls of this experiment:

The conditions used were as follows:
- 1 cycle - 95^oC for 30 seconds
- 30 cycles - 95^oC for 30 seconds, 60^oC for 30 seconds,72^oC for 30 seconds
- 1 cycle - 72^oC for 2 minutes
The numbers of the ligations correspond to the following ligation reactions:
- 7-SacB-E+AK3
- 8-LipA-E+AK3
- 9-P43-gsiB+AK3
- 10-P43-spoVG+AK3
- 11-Pveg-gsiB+AK3
Of the SCP the following look to have given a positive result:
- 7_1,2,3
- 8₁
- 9_1,2
- 10₃
- 11-All too big
11 has failed and in addition there is a contamination within the negativ control. The next step is to repeat the SCP of the contruct 11

@@ Line 9: / Line 9: @@
 ==Wet Lab==
 ===Miniprep===
 *9 minipreps were carried out for samples 5-1, 5-2, 8-1, 8-2, 8-3, 9-2, 9-3, 12-2 and 12-3.
+*Minipreps 8-1, 8-2 and 8-3 digested with ''Eco''RI and ''Spe''I
+[[Image:digest-22.png|center|300px]]
+*Lane 1 - Undigested miniprep 8-1, lane 2 - digested miniprep 8-1, lane 3 - undigested miniprep 8-2, lane 4 - digested miniprep 8-2, lane 5 - undigested miniprep 8-3 and lane 6 - digested miniprep 8-3.
+*The vector in mini-preps 8-1 and 8-2 appear to be ok (~3kb (pSB1AK3)), although there no insert is visible (this may be due to it being too small however). The vector and possible insert in midiprep 8-3 appear to be the wrong size.
 ===Single Colony PCR===