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Team:Wageningen UR/Journal/week24
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''11 October'' | ''11 October'' | ||
| - | * grow 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 | + | * 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 | -> 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 | * 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'' | ''12 October'' | ||
| - | * 2nd time growing 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 | + | * 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 | -> again there is growth on the negative control | ||
-> no green fluorescence could be seen | -> 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 | ||
| + | |||
| Line 25: | Line 53: | ||
---- | ---- | ||
| - | '' | + | ''15 October'' |
* PCR check of the PCR fragment obtained on 27.August | * PCR check of the PCR fragment obtained on 27.August | ||
[[File:PCR check Hepinsidecoil 11.10.png|500px|center|thumb|<p align="justify">''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.'</p>]] | [[File:PCR check Hepinsidecoil 11.10.png|500px|center|thumb|<p align="justify">''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.'</p>]] | ||
| + | |||
| + | * ligation of the PCR product in pJET | ||
| + | |||
| + | * transformation with DH5α | ||
| + | -> there was growth on the negative control plate | ||
| + | |||
---- | ---- | ||
[[https://2012.igem.org/Team:Wageningen_UR/Journal/week23 previous week]] [[https://2012.igem.org/Team:Wageningen_UR/Journal/week25 next week]] | [[https://2012.igem.org/Team:Wageningen_UR/Journal/week23 previous week]] [[https://2012.igem.org/Team:Wageningen_UR/Journal/week25 next week]] | ||
Revision as of 14:11, 22 October 2012
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
