Team:Wageningen UR/Journal/week24

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-> 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  
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''15 October''
 
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* 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
 
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-> no GFP production could be seen
 
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''16 October''
 
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* plate the JM109 samples containing GFPcoil with an IPTG induced promoter and GFPcoil+his tag with an IPTG induced promoter on agarplates containing IPTG
 
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-> culture containing BBa_K883702 shows green fluorescence
 
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* grow colonies containing the biobricks BBa_K883702 BBa_K883703 BBa_K883701 and BBa_K883700
 
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* 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 
 
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* miniprep of the colonies containing BBa_K883702 BBa_K883703 BBa_K883701 and BBa_K883700
 
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* 2nd try to send the bricks BBa_K883702 BBa_K883703 BBa_K883701 and BBa_K883700 in for sequencing
 
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-> sequencing revealed that BBa_K883702 and BBa_K883703 had the expected sequence but BBa_K883701 and BBa_K883700 where faulty
 
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''18 October''
 
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* transformation of BBa_K883702 into BL21 (producing strain) - plating the transformants on plates containing IPTG
 
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'''Hepatitis B inside modification'''
 
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''15 October''
 
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* PCR check of the PCR fragment obtained on 27.August
 
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[[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>]]
 
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* ligation of the PCR product in pJET
 
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* transformation with DH5α
 
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-> there was growth on the negative control plate
 
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[[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 14:33, 23 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


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