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Reporter: Week 8 July 3-July 9
From 2011.igem.org
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
