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From 2012.igem.org

week 17: 20 august - 26 august

Office work

Lab work

Hepatitis B

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20 August

• Colony PCR

- of HepB (without promoter) in BBa_J04450 - of HepB with a his tag + IPTG promoter in Bba_J04500

• 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

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

Hepatitis B inside modification

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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

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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

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