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RecA: Week 3, May 30-June 3
From 2011.igem.org
Contents |
Monday
RecAI Extraction, Day 1
The RecA group extracted the RecAI gene from Escherichia coli using the genomic DNA extraction kit produced by Omega Bio-Tek. The RecA group created a culture of the RecAI gene and left this culture to grow overnight.
Tuesday
RecAI Extraction, Day 2
The RecA group performed miniprep on the RecAI culture from 5/30/11. The RecA group placed the RecAI gene into the C3 vector through amplified insert assembly in order to perform mutagenesis on the RecAI gene. The RecAI team also developed a plan for RecA mutagenesis. Figure 1 shows the overview of this plan. The following table describes each part in more detail.
| Part | Registry Name | Resistance | Position on Registry Plate | Length (bp) | Digested as (insert/vector) |
|---|---|---|---|---|---|
| 1: Terminator | B0015 | A/K | Plate 1 Well 23L | 129 | Vector |
| 2: Reporter (cI/RFP) | I763007 | A/A | Plate 1 Well 15J | 918 | Insert |
| 3: Constitutive Promoter | J23118 | A/A | Plate 1 Well 22A | 35 | Vector |
| 4: cI Repressor | K081007 | A/A | Plate 2 Well 20J | 796 | Insert |
The RecA group plans to insert RecA mutants downstream of the cI Repressor so that the constitutive promoter will continuously transcribe the cI Repressor and the RecA mutant. If the mutant has recombinant activity then the reporter will be deleted because of the homologous sequences of the Terminator (Part 1) and the terminator of the Reporter due to homologous sequences of the Terminator (Part 1) and the terminator of the Reporter, part 2 will be deleted.
