Team:Wageningen UR/Journal/week26
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+ | |||
+ | ''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 | ||
+ | |||
+ | [[File:Fluorescent microscope JM109.png|500px|center|thumb|<p align="justify">''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) '</p>]] | ||
+ | |||
+ | |||
+ | ---- | ||
+ | [[https://2012.igem.org/Team:Wageningen_UR/Journal/week25 previous week]] | ||
Revision as of 09:54, 26 October 2012
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