Team:Newcastle/6 July 2010

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(New page: '''Gram-positive Bacteria Chromosomal DNA Extraction Protocol''' ===Puregene DNA isolation=== # Grow overnight 7.5ml cultures in THYB containing 30μg/ml hyaluronidase. ===Cell L...)
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'''[[Gram-positive Bacteria Chromosomal DNA Extraction Protocol]]'''
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{{Team:Newcastle/mainbanner}}
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===Puregene DNA isolation===
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# Grow overnight 7.5ml cultures in THYB containing 30μg/ml hyaluronidase.
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===Cell Lysis===
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=Genomic DNA extraction from ''Bacillus subtilis'' strain ATCC 6633=
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# Cells go through 3600 x g centrifugation for 15 minutes to pellet.  
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# The supernatant is poured away.
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==Aim==
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# 0.5ml of cell suspension solution is added to the eppendorf tube. The solution in the tube is pipetted up and down to resuspend cells. The solution is then transferred to another 1.5ml tube.
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The aim of this experiment was to extract genomic DNA from ''Bacillus subtilis'' strain ATCC 6633. This strain has the subtilin genes, required for our Subtilin Production and Immunity parts.
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# 50μl of lysozyme and 7.5μl of lytic enzyme solution is added into the tube. The tube is then inverted 25 times to mix the solution.
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# We then incubate it at 37°C
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==Protocol==
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Please refer to: [[Team:Newcastle/DNA extraction|DNA extraction of ''Bacillus subtilis'']] for materials required and protocol.
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==Results==
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Results were as expected
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see: https://2010.igem.org/Team:Newcastle/7_July_2010
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===Conclusion===
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See: https://2010.igem.org/Team:Newcastle/7_July_2010
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=Research=
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Today we also continued with our research on filamentous cells which will act as reinforcing fibres in the crack.
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#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.
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#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.  
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{{Team:Newcastle/footer}}

Latest revision as of 15:08, 27 October 2010

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Contents

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 has the subtilin genes, required for our Subtilin Production and Immunity parts.


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.


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