Team:Grenoble/Biology/Notebook/July/week 30

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July

week 27 week 28 week 29 week 30

Week 30: July 23rd to 29th

Goal of the week:

Test the Gibson Assembly and the transformation protocols, recover and amplify some biobricks involved in our genetic networks :
  • pAra/Bad_RBS_GFP (1300bp)
  • RBS_Cya (2600bp)
  • fha (80bp)
  • eCFP (800bp)
  • pSB4K5 (2400bp)

Monday, July 23rd:

We did PCRs with HF Phusion enzyme (protocol) on miniprep in order to amplify pAra/Bad_RBS_GFP, and on colony to amplify RBS_Cya. We did the PCR both with and without DMSO.

To separate the PCR products (12/07/23 and 12/07/20), we prepared a 1.8% TAE agarose gel.
Migration conditions = 100V during 30 min.
In order to reveal the DNA fragments, we used EtBr.

photo_gel_16
Migration result for a 1.3% TAE agarose gel
(the DNA ladder scale is in kb)
Lane 1: DNA ladder 80pb-10kb (fermentas)
Lane 2: pSB4K5 PCR product (DMSO)
Lane 3: fha1 PCR product (DMSO)
Lane 4: fha1 PCR product
Lane 5: pSB4K5 PCR product
Lane 6: pAra/Bad_RBS_GFP PCR product (DMSO)
Lane 7: pAra/Bad_RBS_GFP PCR product (DMSO)
Lane 8: RBS_Cya PCR product (DMSO)
Lane 9: RBS_Cya PCR product
Lane 10: pAra/Bad_RBS_GFP PCR product
Lane 11: pAra/Bad_RBS_GFP PCR product
Lane 12: DNA ladder 80bp-10kb (fermentas)

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