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| - | {{EPF-Lausanne09}}
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| - | <div CLASS="epfltrick">__TOC__
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| - | </div><div CLASS="epfl09">
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| | | | |
| - | =Wet Lab=
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| - | ==July==
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| - | ===06.07.09===
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| - | LOVTAP plasmid AND TrpR plasmid were transformed in competent E. Coli following received protocol, and grown overnight (see Lab book for more details).
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| - | <br>One problem: we actually don't know TrpR plasmid resistance, so we tried with three resistances available in the lab: Amp., Kana. and Chl.
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| - | LOVTAP is in a plasmid called pCal-n (see picture below):
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| - | [[Image: pCAL-n.jpg|500px|thumb|center|pCal-n plasmid]]
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| - | <br>Some comments on the plasmid:
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| - | <br>-CBP is a small peptide with which we could purify LOVTAP protein
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| - | <br>-Thrombin target is a nucleotidic sequence that can be recognized by thrombin a peptidase. This peptidase will cut CBP once LOVTAP is purified
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| - |
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| - | ===07.07.09===
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| - | We have to grow the 3 strains generously sent by [mailto:j.beatty@ubc.ca Tom Beatty]
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| - | The three strains are :
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| - | :*''R.Palustris'' CEA001 (wild type) ; should be grown on LB medium only
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| - | :*''R.Palustris'' BPHP1+ ; should be grown on LB with gentamycin (100 micrograms/ml)
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| - | :*''E.Coli'' DH10B (pBPH/hmu0) ; should be grown on LB with gentamycin (20 micorgrams/ml)
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| - | The transformed LOVTAP and TrpR worked well (N.B. the plasmid of TrpR is pUC19 so the antibiotic resistance is Amp -> see below)
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| - |
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| - | [[Image: RTEmagicC_puc19_2.gif.gif|500px|thumb|center|pUC19 plasmid]]
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| - | We did the glycerol stock, located in the -80 fridge, first floor of the iGEM compartement.
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| - |
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| - | Then, a miniprep was done with both cultures.
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| - | A LOVTAP plasmid aliquot was done, a TrpR plasmid aliquot was done, located in the -20 fridge, 2nd floor.
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| - |
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| - | ===08.07.09===
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| - | 1. R. Palustris culture grew. A glycerol stock has been done.
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| - | A pellet is on the fridge level 2, waiting for a miniprep.
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| - | 2. iGEM parts have been transformed:
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| - | <br>- BBa_B0010 (terminator), resistance: A, well kit plate 1: 13D
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| - | <br>- BBa_R0010 (promoter LacI), resistance: A, well kit plate 1: 1D
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| - | <br>- BBa_B0030 (RBS), resistance: A, well kit plate 1: 1H
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| - | <br>- BBa_E0240 (RBS-GFP-TER), resistance: A, well kit plate 1: 12M
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| - | <br>- BBa_I13507(RBS-mRFP-TER), resistance: A, well kit plate 1: 22O
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| - | <br>- BBa_J13002(pTetR-RBS), resistance: A, well kit plate 1: 13B
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| - | <br>- BBa_I6007 (inverter TetR), resistance: A, well kit plate 2: 1C
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| - |
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| - | {| class="wikitable" width="80%" align="center"
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| - | |+ '''Parts&Characteristics'''
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| - | |-
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| - | ! scope=col | Part
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| - | ! scope=col | Resistance
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| - | ! scope=col | Well (Kit Plate)
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| - | |-
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| - | | width="33%" align="center" |
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| - | BBa_B0010 (terminator)
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| - | | width="34%" align="center" |
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| - | A
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| - | | width="34%" align="center" |
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| - | 13D (1)
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| - | |-
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| - | | width="33%" |
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| - | BBa_R0010 (promoter LacI)
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| - | | width="34%" |
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| - | A
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| - | | width="34%" |
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| - | 1D (1)
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| - | |-| align="center" colspan="3" |
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| - | Ligne 1
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| - | |}
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| - |
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| - | ===09.07.09===
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| - |
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| - | ===10.07.09===
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| - |
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| - | ==August==
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| - |
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| - | </div><div CLASS="epfl09bouchon"></div>
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