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Team:Wageningen UR/Journal/week26
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Week 26: 22 october - 28 october
GFP modification
22 October
- sent bricks BBa_K883704 and BBa_K883705 in for sequencing (single read using the reverse sequencing primer)
-> the bricks have the expected sequence
- seperated colonies containing BBa_K883700 and BBa_K883701 (GFPcoil and GFPcoil with a His tag in pSB1C3) from the plate of 21.october a second time
23 October
- colony PCR on the seperated colonies containing the faulty biobricks BBa_K883700 and BBa_K883701 to check wheather there are colonies present with the correct insert
-> there was no insert present in the picked colonies
- grow batches BBa_K883702 in BL21, BBa_K883703 in JM109, BBa_K883704 in DH5α, BBa_K883705 in DH5α
24 October
- plate BBa_K883702 in BL21, BBa_K883703 in JM109, BBa_K883704 in DH5α, BBa_K883705 in DH5α on plates containing IPTG together with a control that does not produce fluorescence
- checked fluorescence of JM109 cultures (from a plate) containing BBa_K883702 and BBa_K883703 with fluorescence microscopy
-> fluorescence could be seen in both cultures; a negative control (JM109 without a GFP encoding plasmid) showed no fluorescence; the positive control (JM109 containing Bba_I13522 - GFP with a constitutive promoter) showed strong fluorescence
Figure 1: fluorescence microscopy pictures of a negative control (JM109 without a GFP encoding plasmid); JM109 cultures containing the bricks BBa_K883702 GFP-coil and BBa_K883703 GFP-coil with His tag and a positive control (JM109 containing Bba_I13522 - GFP with a constitutive promoter) '
