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Team:Wageningen UR/Journal/week24
From 2012.igem.org
Week 24: 8 october - 14 october
GFP modification
11 October
- grow JM109 containing our biobricks BBa_K883702 (GFP coil with an IPTG induced promoter) and BBa_K883703 (GFP coil + His tag with an IPTG induced promoter)and induce expression with IPTG
-> no conclusion could be made due to growth on the negative control
- send samples of BBa_K883702 BBa_K883703 BBa_K883701 and BBa_K883700 in for sequencing to check if the last transformation from pJET into the iGEM backbones succeded
-> sequencing results where never obtained - the samples did not arrive
12 October
- 2nd time growing JM109 containing our biobricks BBa_K883702 (GFP coil with an IPTG induced promoter) and BBa_K883703 (GFP coil + His tag with an IPTG induced promoter) and induce expression with IPTG
-> again there is growth on the negative control -> no green fluorescence could be seen
15 October
- 3rd time growing JM109 containing our biobricks BBa_K883702 (GFP coil with an IPTG induced promoter) and BBa_K883703 (GFP coil + His tag with an IPTG induced promoter) and induce expression with IPTG with new medium
-> no GFP production could be seen
16 October
- plate the JM109 samples containing GFPcoil with an IPTG induced promoter and GFPcoil+his tag with an IPTG induced promoter on agarplates containing IPTG
-> culture containing BBa_K883702 shows green fluorescence
- grow colonies containing the biobricks BBa_K883702 BBa_K883703 BBa_K883701 and BBa_K883700
- grow colonies containing BBa_K883702 and BBa_K883703 in duplo once with IPTG in the medium and once adding IPTG (fresh stock solution) to the culture at OD=0.6
- miniprep of the colonies containing BBa_K883702 BBa_K883703 BBa_K883701 and BBa_K883700
- 2nd try to send the bricks BBa_K883702 BBa_K883703 BBa_K883701 and BBa_K883700 in for sequencing
-> sequencing revealed that BBa_K883702 and BBa_K883703 had the expected sequence but BBa_K883701 and BBa_K883700 where faulty
18 October
- transformation of BBa_K883702 into BL21 (producing strain) - plating the transformants on plates containing IPTG
Hepatitis B inside modification
15 October
- PCR check of the PCR fragment obtained on 27.August
Figure 1: The purified PCR fragment contains fragments of the correct size as well as a bigger fragment. The nonpurified sample shows only a smere.'
- ligation of the PCR product in pJET
- transformation with DH5α
-> there was growth on the negative control plate
