Team:Grenoble/Biology/Notebook/July

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<li>Strains = BW25113 WT, BW25113 cya- and BW25113 cya- pAra/Bad </li>
<li>Strains = BW25113 WT, BW25113 cya- and BW25113 cya- pAra/Bad </li>
<li>Conditions = LB liquid medium, 37°C, 200rpm, overnight </li>
<li>Conditions = LB liquid medium, 37°C, 200rpm, overnight </li>
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<h2>Wednesday, July 04<span class="exposant">th</span>:</h2>
<h2>Wednesday, July 04<span class="exposant">th</span>:</h2>
We did PCRs on colony with a GoTaq enzyme (protocol) to amplify:
We did PCRs on colony with a GoTaq enzyme (protocol) to amplify:

Revision as of 13:22, 6 August 2012

iGEM Grenoble 2012

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iGEM 2012
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week 27 week 28 week 29 week 30

Week 27: July 02nd to July 08st

Goal of the week

We wanted to test the PCR conditions, recover and amplify the biobricks involved in our genetic networks :
  1. pAra/Bad_RBS_GFP (1300bp)
  2. RBS_Cya (2600bp)
  3. pLAC (100bp)
  4. fha (80bp)
  5. eCFP (800bp)
  6. pLAC_RBS (120bp)
  7. RsmA (200bp)
  8. rsmY (170bp)
  9. pSB1A3 (2400bp)
  10. pSB4K5 (2400bp)
  11. pSB3C5 (2400bp)

Tuesday, July 03rd:

Precultured cells are prepared:
  1. Strains = BW25113 WT, BW25113 cya- and BW25113 cya- pAra/Bad
  2. Conditions = LB liquid medium, 37°C, 200rpm, overnight

Wednesday, July 04th:

We did PCRs on colony with a GoTaq enzyme (protocol) to amplify:
  1. fha1, RsmA, rsmY and pSB1A3 from iGEM Grenoble 2011 glycerol stock.
  2. pAra/Bad_RBS_GFP from BW25113 cya- pAra/Bad precultured cells.
  3. RBS_Cya from BW25113 WT precultured cells.

Using iGEM 2012 biobricks we transformed BW25113 WT cells (protocol). We obtained four transformed strains with four different biobricks:
  1. BBa_I13601: pLAC_RBS
  2. BBa_E0422: eCFP
  3. pSB3C5 plasmid
  4. psB4K5 plasmid

Thursday, July 5th:

The transformations (12/07/04) show no growth on LB Agar plate + antibiotics.

In order to separate the PCR products (12/07/04), we prepared two gels:
  1. a 1.3% Tris base Acetic Acid EDTA (TAE) agarose gel, for the small fragments: RsmA, fha1 and rsmY (length < 1000bp)
  2. a 0.8% TAE agarose gel, for the big fragments: pSB1A3, RBS_Cya, and pAra/Bad_RBS_GFP (length > 1000bp)

    3. Migration conditions = 100V during 35 min. In order to reveal the fragments, we used Ethidium Bromide (EtBr).

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