Newcastle University Wetlab/17 September 2008

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==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