Revision as of 17:37, 5 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]

4 September 2008

Wetlab

Cloning

Pfu PCR reactions for LacI gene, Aad9 gene and AmyE 3' Integration Sequence set up using the standard conditions and reagents [http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Wetlab/PCR]; LacI and AmyE 3' at 52°C for the first 10 cycles and 65°C for the last 20, Aad9 at 52°C then 60°C. 5μl of each PCR reaction run on a gel - results below
CAT biobrick digested with EcoRI and SpeI ready for assembly with the terminator
Termiantor biobrick cut with EcoRI and XbaI ready for assembly with CAT
GFP-Termiantor biobrick cut again with XbaI and SpeI to produce biobrick vectors for our PCR clones
GFP-Termiantor biobrick cut again with EcoRI and SpeI to produce biobrick vectors for our GeneArt produced clones

Preparation of Electrocompetent E. coli Cells

We have prepared 42 aliquots of electrocompetent XL1 Blue E. coli cells using [http://openwetware.org/wiki/IGEM:IMPERIAL/2008/New/Protocols/XL1-Blue_preparation the relevant protocol] listed on our Protocols Page. The protocol was updated to include a higher innoculation value from the overnight culture.

Results

A 1% Agarose gel showing the results of various PCR reactions. Each lane is loaded with 5ul of PCR reaction and 1ul of 6x sample buffer. M = Marker, lane 1 is the AmyE 3' Integration sequence, lane 2 the LacI and lane 3 the Aad9 PCR reaction

As can be seen, both the AmyE 3' integration sequence and the Aad9 (spectinomycin resistance) gene amplified properly. LacI did not however and so this will need to be repeated.

@@ Line 24: / Line 24: @@
 ===Results===
-[[Image:4-08PCR.PNG|thumb|600px|center|A 1% Agarose gel showing the results of various PCR reactions. Each lane is loaded with 5ul of PCR reaction and 1ul of 6x sample buffer. M = Marker, lane 1 is the AmyE 3' Integration sequence, lane 2 the LacI and lane 3 the Aad9 PCR reaction]]
+[[Image:4-08PCR.PNG|thumb|center|A 1% Agarose gel showing the results of various PCR reactions. Each lane is loaded with 5ul of PCR reaction and 1ul of 6x sample buffer. M = Marker, lane 1 is the AmyE 3' Integration sequence, lane 2 the LacI and lane 3 the Aad9 PCR reaction]]
 As can be seen, both the AmyE 3' integration sequence and the Aad9 (spectinomycin resistance) gene amplified properly. LacI did not however and so this will need to be repeated.

Imperial College/4 September 2008

From 2008.igem.org