Team:TUDelft/19 August 2009

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19 August 2009

Tim Weenink

After yesterdays horrible discovery that the I-SceI Homing Endonuclease we got from MIT was actually T4 DNA ligase we had to adjust our cloning strategy. The more primitive part without promoter was found to be the correct one, except that is was assembled with part !A instead of p(LacI). So we did that assembly again today. With p(LacI) from the delay team upstream, *T1 (downstream) and backbone pSB1C3 (also from the delay team). Both RbCl chemically comp and EC electrocomp cells were transformed.


Saeed

Saeed19082009wiki.jpg

lane Part Expected Plasmid Size Status
1 [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder]
2  !Fr1n upstream 1900 ok, but not needed
3  !Fr1n downstream 1900 ok
4 destination plasmid 3000 ok

!Fr1n downstream will be purified from the gel by gel extraction and used for the new stategy as Tim mentioned before.

Daniel

Digestion of biobricks:

Gel180809.jpg

Well Biobrick Expected Plasmid Size Status
1 [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder]
2 C0051 750
3 I13507 861
4 P0440 840
5 I13504 875
6 J04630 857
7 R0051 44 too small
8 B0015 129
9 J13002 74
10 pSB1C3 2072
11 pSB1AK3 3426
12 pSB1A3 2157
13 CB
14 CC
15 [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder]

Assemblies 1A, 2A, 3A, 4A, 5A and 6A

Transformation and culture in plates and tubes