Genomic DNA extraction from Bacillus subtilis strain ATCC 6633

Aim

The aim of this experiment was to extract genomic DNA from Bacillus subtilis strain ATCC 6633. This strain produces the subtilin genes, required for our Subtilin Production and Immunity BioBricks as well as the Quorum sensor biobrick designed by Team Newcastle 2008.

Materials and methods

• ara forward and reverse primers
• sac forward and reverse primers
• Pipettes
• Universal tubes
• Eppendorf tubes
• Centrifuge
• PCR (see PCR protocol from Lab training)
• Gel electrophoresis (see Gel electrophoresis from Lab training)

ara and sac are two genes that exist in the ATCC 6633 strain. ara and sac forward and reverse primers are two tests, which will be used in PCR. At the end, Gel Electrophersis can be used to distinguish the bands. If the bands from either of ara or sac show up at the expected bps against the DNA ladder, this proves that the genomic DNA is sucessfully extracted.

Protocol

Please refer to: DNA extraction of Bacillus subtilis for materials required and protocol.

Results

Results were as expected see: https://2010.igem.org/Team:Newcastle/7_July_2010

Conclusion

See: https://2010.igem.org/Team:Newcastle/7_July_2010

Research

Today we also continued with our research on filamentous cells which will act as reinforcing fibres in the crack.

1. Baarle S van and Bramkamp M. (2010). "The MinCDJ system in Bacillus subtilis prevents minicell formation by promoting divisome disassembly". PloS one. 5(3). e9850.
2. Mo AH and Burkholder WF.(2010). "YneA, an SOS-induced inhibitor of cell division in Bacillus subtilis, is regulated posttranslationally and requires the transmembrane region for activity". Journal of bacteriology. 192(12). 3159-73.