Team:Wageningen UR/Journal/week17

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(Difference between revisions)
(→week 17: 20 august - 26 august)
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== Lab work ==
== Lab work ==
 +
 +
'''Hepatitis B'''
'''Hepatitis B'''
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- Of the transformations HepB + IPTGpromoter in BBa_J04450 backbone in JM109 and HepB + IPTGpromoter in BBa_J04450 backbone in DH5α
- 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'''
'''Hepatitis B inside modification'''
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* control of a miniprep of HepB+IPTG in BL21 from 16.Aug (sample from 6.Aug)
* 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
-> 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
 +
 +
[[File:PCR 2nd,3rd step hepinsidecoil.png|200px]]
 +
 +
-> 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'''
'''GFP modification'''
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 +
''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
----
----
[[https://2012.igem.org/Team:Wageningen_UR/Journal/week16 previous week]]          [[https://2012.igem.org/Team:Wageningen_UR/Journal/week18 next week]]
[[https://2012.igem.org/Team:Wageningen_UR/Journal/week16 previous week]]          [[https://2012.igem.org/Team:Wageningen_UR/Journal/week18 next week]]

Revision as of 14:17, 17 September 2012

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

  • 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

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

PCR 2nd,3rd step hepinsidecoil.png

-> 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

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