EPF-Lausanne/8 July 2009

From 2009.igem.org

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===Wet Lab===
===Wet Lab===
1. R. Palustris culture grew. A glycerol stock has been done.
1. R. Palustris culture grew. A glycerol stock has been done.
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:LOVTAP is digested with ES and inserted into Term plasmid (which was digested with EX previously).
:LOVTAP is digested with ES and inserted into Term plasmid (which was digested with EX previously).
:Finally, LacI promoter-RBS digested with ES to be inserted into LOVTAP-Term plasmid, digested with EX.
:Finally, LacI promoter-RBS digested with ES to be inserted into LOVTAP-Term plasmid, digested with EX.
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==People in the lab==
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: Heidi, Tu, Nath, Rafael, Basile
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Revision as of 15:42, 27 July 2009



Contents

Wet Lab

1. R. Palustris culture grew. A glycerol stock has been done. A pellet is on the fridge level 2, waiting for a miniprep.

2. iGEM parts have been transformed:

Parts&Characteristics
Part Characteristic Resistance Well (Kit Plate)

[http://partsregistry.org/Part:BBa_B0010 BBa_B0010]

Terminator

A

13D (1)

[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]

Promoter LacI

A

1D (1)

[http://partsregistry.org/Part:BBa_B0030 BBa_B0030]

RBS

A

1H (1)

[http://partsregistry.org/Part:BBa_E0240 BBa_E0240]

RBS-GFP-TER

A

12M (1)

[http://partsregistry.org/Part:BBa_I13507 BBa_I13507]

RBS-mRFP-TER

A

22O (1)

[http://partsregistry.org/Part:BBa_J13002 BBa_J13002]

pTetR-RBS

A

13B (1)


Cloning Strategy

Inducible LOVTAP biobrick strategy

  • Problem to overcome:
PstI sites in LOVTAP sequence.
  • Our goal:
Biobrick consisted of LacI promoter-RBS-LOVTAP-Term (in this order).
  • Material:
Biobrick of LacI promoter, RBS, LOVTAP, Term separately ( LOVTAP obtained from previous section and the rest from iGEM Spring 2009 distribution Kit plate).
  • Strategy:
LacI promoter-RBS ligation with iGEM protocol (LacI promoter digested with ES, RBS digested with XP, plasmid containing an other antibiotic digested with EP).
LOVTAP is digested with ES and inserted into Term plasmid (which was digested with EX previously).
Finally, LacI promoter-RBS digested with ES to be inserted into LOVTAP-Term plasmid, digested with EX.


People in the lab

Heidi, Tu, Nath, Rafael, Basile
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