Newcastle University Wetlab/18 September 2008

Thursday 18th September



 * Plasmid was isolated from 2mL of each of the 6 ON cultures. pGFPrrnB-ncl08 colonies 7 + 11 were restricted using 7.5μl plasmid and 0.5μl each enzyme (EcoRI and NheI) in 15μl total volume.

Lane 1: 1 kb ladder Lane 2: pGFPrrnB-ncl08 colony 7 restricted with EcoRI + NheI Lane 3: pGFPrrnB-ncl08 colony 11 restricted with EcoRI + NheI Lane 4: -

This showed that glycerol stocks of these plasmids are good as the fragment sizes are as expected. pGFPrrnBrrnB colony 7 is particularly strong and contains a large quantity of DNA.


 * iGEMgfp colony 7.1 was induced with subtilin and prepared for microscopy. This showed no difference between the subtilin-induced and the control cultures. Also, only some cells expressed gfp whilst others did not. However, it was noticed that every cell that did flouresce was also undergoing sporulation, possibly indicating that the insert promotor may be under the control of the sporulation factor.


 * ON agar plates of iGEMBB107 colony 3 and 5 (plated on starch + chloramphenicol, spectinomycin and chloramphenicol) were analysed. Colonies were wanted that did not have a 'halo' on starch + cam, that did not grow on spec and that did grow on cam. All but colony 3.9 showed to be good. Therefore ON cultures were made from the cam only plates of colonies 3.1, 3.2, 5.1 and 5.2 in 3mL LB.


 * ON culture was also made from iGEMcherry colony 13.1 glycerol stock in 3mL LB for microscopy.