Team:Newcastle/26 July 2010

From 2010.igem.org

Revision as of 19:40, 26 July 2010 by Swoodhouse (Talk | contribs)

iGEM Homepage Newcastle University BacillaFilla Homepage Image Map

Contents

Preparation for cloning of the rocF BioBrick

A

  • Re-hydrated [http://partsregistry.org/Part:pSB1C3 pSB1C3] (the plasmid we will be submitting our BioBricks to the registry in) and [http://partsregistry.org/Part:BBa_B0014 BBa_B0014] (the double terminator we will be using for the rocF BioBrick) from the parts distribution.
  • Transformed and plated E. coli DH5α

Overnight cultures of B. subtilis 168 for chromosomal DNA extraction

Colony PCR of Genomic DNA

Aim:

To determine whether the genes have been inserted into the plasmid of B. Subtilis 168.

Materials:

  • Pipette
  • Microfuge
  • Microtubes
  • Distilled H2O
  • Nucleotide DNTP
  • 5x GoTaq buffer
  • Template DNA (B. Subtilis ATCC 6633, 1:1 and 1:2)
  • Forward and reverse primers

Protocol:

  • For the full protocol, please refer to Colony PCR in Protocol List.

Conditions in ThermoCycler:

  • Melting temperature, Tm used for Anneal step is 59°C.

Results:

Gel electrophoresis will be run tomorrow to determine the results.

Conclusion:

Please refer to Lab book dated 27.7.2010.

Newcastle University logo.png    Newcastle cbcb logo.pngNewcastle Biomedicine logo.gif    Team Newcastle CEG logo.gif
Newcastle iww logo.jpg  UNIPV Pavia Logo.gif  Newcastle BBSRC.gif    Newcastle Genevision logo.png Newcastle WelcomeTrust.jpg
FaceBook Icon