Reporter: Week 8 July 3-July 9

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Contents

Sunday, July 3

Assembly of Fusion Parts, Day 5

     All of the assemblies from 7/2 and the promoter+RBS assembly (J23100+B0034) were verified using agarose gel electrophoresis. Since the ladder was contaminated, the relative sizes of the assemblies were analyzed. This analysis showed that the J23100+B0034 was much smaller than the other assemblies, the XylE+both linkers were just under 1000 base pairs, and the GFP+cleavage sites were a little smaller than the XylE+linker parts. Thus, each construct was grown in culture overnight in order to extract plasmids for sequencing.

     Plasmids from the four cultures made on 7/1 were extracted using the Omega Bio-Tek miniprep protocol. These plasmids were stored in the 4°C fridge until they could be sent to sequencing on 7/5.

Monday, July 4

Assembly of Fusion Parts, Day 6

     Plasmids were extracted from the eight cultures made on 7/3 using the Omega Bio-Tek miniprep protocol. Since the sequencing center is closed for the holiday, these plasmids will be sent to sequencing tomorrow, along with the plasmids extracted on 7/2.

Tuesday, July 5

The following plasmids were sent to sequencing:

J23100+B0034 (A and B)

LacZ+Imp linker

LacZ+Small linker

LacZ+10 AA linker

XylE+Imp linker

XylE+Small linker

XylE+10AA linker

GFP+cI cleavage site (A and B)

GFP+tev cleavage site (A and B)

Results: The J23100+B0034 (colony A) sequence results showed that the


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