Team:Newcastle/28 July 2010

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Contents

PCR Experiment

Aims

The aim of this experiment is to prove that Bacillus subtilis 168 and 3610 DNA extraction worked by amplifying anaR gene using PCR.

Materials and Protocol

Please refer to: PCR for GoTaq PCR.

Result

  • Lane 1: 1kb DNA ladder
  • Lane 2: B. subtilis 168 chromosomal DNA containing anaR gene
  • Lane 3: B. subtilis 168 chromosomal DNA containing anaR gene
  • Lane 4: B. subtilis 3610 chromosomal DNA containing anaR gene
  • Lane 5: B. subtilis 3610 chromosomal DNA containing anaR gene

Discussion

We found bands in the lane 2, 3, 4 and 5 of around 200 bp size which is an approximate size of the anaR gene which is found on the chromosome of both B. subtilis 168 and 3610.

Conclusion

This experiment proves that the DNA extraction from both B. subtilis 168 and 3610 done on 27th July, 2010 was successful.

Plasmid Miniprep Experiment

Aims

The aim of this experiment is to extract plasmid DNA pSB1C3, pSB1AK3 and plasmid containing lacI Biobrick from E. coli DH5α cells with the help of Qiagen miniprep kit and confirming the extraction with the help of nanodrop experiment.

Materials and Protocol

Please refer to: Minipreps for Qiagen miniprep protocol and Nanodrop Spectrophotometer for nanodrop protocol.

Result

  • Lane 1: 1kb DNA ladder
  • Lane 2: Extraction of pSB1C3 plasmid
  • Lane 3: Extraction of pSB1C3 plasmid
  • Lane 4: Extraction of plasmid containing lacI
  • Lane 5: Extraction of plasmid containing lacI
  • Lane 6: Extraction of pSB1AK3 plasmid containing double terminator
  • Lane 7: Extraction of pSB1AK3 plasmid containing double terminator

Discussion

We found bands in the lane 2, 3, 4 and 5 of around 200 bp size which is an approximate size of the anaR gene which is found on the chromosome of both B. subtilis 168 and 3610.

Conclusion

This experiment proves that the DNA extraction from both B. subtilis 168 and 3610 done on 27th July, 2010 was successful.


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