Sensor: Week 7 June 28-July 1
From 2011.igem.org
Contents |
Tuesday, June 28
cI Repressor Test Assembly, Day 23
The sensor group continued their assembly of their testing construct. They placed an RBS in the K3 vector with Cro, the OR in front of the mCherry reporter, terminator, cI repressor and J23113 promoter as well as in front of the 34mCherry. Finally, the sensor group attempted (again) to put the B0032 RBS in front of the cI repressor.
Protocol | Part Involved with Protocol | |
---|---|---|
Restriction Digest | Inserts using XbaI and PstI: | 34mCherry K648000+K648001 |
Inserts using EcoRI and SpeI: | B0032 B0034 | |
Vectors using EcoRI and XbaI: | C0051 K3+Cro | |
Vector using SpeI and PstI: | K3+OR | |
Ligation | B0034 + K3 + Cro KOR + K648000 + K648001 KOR + 34mcherry B0032 + C0051 | |
Transformation/Plating | The above ligations were transformed into Escherichia coli cells and plated on kanamycin resistant plates. |
The sensor group also transformed K3+Cro and J23113+B0031+C0051 in order to make stocks. The B0034+C0051 was grown up from stock.
Wednesday, June 29
cI Repressor Test Assembly, Day 24
The sensor group verified their assemblies from 6/28. Colonies containing KOR + 34mcherry, B0034 + K3 + Cro, KOr + K648000+K648001 were amplified through colony PCR.
The B0034 and B0034+C0051 plasmids were extracted using the Omega-Biotek miniprep protocol.