Newcastle University Wetlab/17 September 2008

Wednesday 17th September

 * Results of ON agar plate cultures:

- pGFPrrnB-BBa_I746107 colony 3 (chloramphenicol) = ~100 colonies

- pGFPrrnB-BBa_I746107 colony 5 (chloramphenicol) = ~80 colonies

- B. subtilis 168 (negative control) (chloamphenicol) = O colonies

These colonies show that the BBa_I746107 insert has successfully transformed into the B. subtilis chromosomal DNA. To avoid confusion between E. coli and B. subtilis, it was decided to rename the insert from pGFPrrnB-ncl08 (in E. coli) to iGEMBB107 (in B. subtilis). It was important to then check that only this insert had integrated (and not the rest of the vector plasmid) and that it had integrated in the correct place. Therefore the following agar plate cultures were made:

- iGEMBB107 colony 3 (starch + chloramphenicol)

- iGEMBB107 colony 3 (spectinomycin)

- iGEMBB107 colony 3 (chloramphenicol)

- iGEMBB107 colony 5 (starch + chloramphenicol)

- iGEMBB107 colony 5 (spectinomycin)

- iGEMBB107 colony 5 (chloramphenicol)


 * ON cultures were made from glycerol stocks to check for correct insert (in 3mL LB):

- pGFPrrnB-ncl08 colony 7

- pGFPrrnB-ncl08 colony 11

- iGEMgfp colony 7.1

- iGEMgfp colony 11.1

- iGEMcherry colony 13.1

- iGEMcherry colony 13.2