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Contents 1 Gel electrophoresis for the subtilin immunity fragments 1.1 Aims 1.2 Materials and protocol 1.3 Results 1.4 Discussion

Gel electrophoresis for the subtilin immunity fragments

Aims

The aim of the experiment is to check whether we have the correct fragment sizes on the four subtilin fragments that are we have amplified yesterday.

Materials and protocol

Please refer to the gel electrophoresis and the gel extraction protocols.

Results

Figure 1: Gel electrophoresis of the PCR products of the parts required for the subtilin immunity BioBrick.

• Lane 1: 1 kb DNA ladder
• Lane 2: Plasmid Vector (pSB1C3)
• Lane 3: Promoter and RBS (pVeg-SpoVG)
• Lane 4: spaIFEG Gene Cluster
• Lane 5: Double terminator
• Lane 6: 100 bp DNA ladder

Discussion

Plasmid Vector (lane 2), Promoter & RBS (lane 3) and Double terminator (lane 5) showed up. spaIFEG PCR tube (lane 4) did not show up. We think that this occurrence was due to the Tm error (it should be 63°C as opposed to 46°C). Therefore, another PCR and gel electrophoresis were performed. Please see tomorrow's lab book page for this.

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