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Team:Wageningen UR/Journal/week17
From 2012.igem.org
week 17: 20 august - 26 august
Office work
Lab work
Hepatitis B
20 August
- Colony PCR
- of HepB (without promoter) in BBa_J04450 - of HepB with a his tag + IPTG promoter in Bba_J04500 -> the samples show no band on the gel - so the transformation did not succeed
- Ligation
- of the HepB with the IPTG promoter into BBa_J04450
- Transformation
- Both into JM109 and DH5α; using the BBa_J04450 brick as positive control
22 August
- Colony PCR
- Of the transformations HepB + IPTGpromoter in BBa_J04450 backbone in JM109 and HepB + IPTGpromoter in BBa_J04450 backbone in DH5α -> the samples show no band on the gel - so the transformation did not succeed
23 August
- 2nd try - Colony PCR
Of the transformations GFP-coil in BBa_J04450 backbone in JM109 and GFP-coil in BBa_J04450 backbone in DH5α -> again no band on the gel
Hepatitis B inside modification
23 August
- Step 3 and step 4 of the PCR reaction
-> no bands visible for the 3rd and 4th step of HepBinside-coil
- control of a miniprep of HepB+IPTG in BL21 from 16.Aug (sample from 6.Aug)
-> the miniprep shows a band of the expected size - so the transformation from 6.Aug was successful
24 August
- repeat 3rd step of the PCR reaction with the sample of 15. Aug
-> 3rd step of the PCR reaction seems to work (right size + size difference compared to the 2nd step, but also a smear is visible
GFP modification
20 August
- Ligation
- of the digested GFP-coil PCR product from 15.Aug into BBa_J04450
- Transformation
Both into JM109 and DH5α; using the BBa_J04450 brick as positive control
22 August
- Colony PCR
Of the transformations GFP-coil in BBa_J04450 backbone in JM109 and GFP-coil in BBa_J04450 backbone in DH5α
23 August
- 2nd try - Colony PCR
Of the transformations GFP-coil in BBa_J04450 backbone in JM109 and GFP-coil in BBa_J04450 backbone in DH5α
24 August
- check and amplify GFP-coil PCR product from 14.Aug
-> no conclusion can be made because of very weak bands
