IGEM:Paris Bettencourt 2012/Notebooks/Semantic group/day by day//2012/07/22: Difference between revisions

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* 4°C -<math>\infty</math>
* 4°C -<math>\infty</math>
-> 24 cycles
-> 24 cycles
==Biobricks retrieval==
* Following the [[IGEM:Paris_Bettencourt_2012/Protocols|protocole]], I took BB
{| border = "1" width="45%"
! width="15%" | Parts number
! width="15%" | Name
! width="15%" | Distribution/Plate/Well
! width="15%" | Backbone
|-
|[http://partsregistry.org/Part:BBa_I14034 BBa_I14034]
|P(Kat)
|2012/2/13B
|pSB2K3
|-
|[http://partsregistry.org/Part:BBa_B0034 BBa_B0034]
|RBS
|2012/1/2M
|pSB1A2
|-
|[http://partsregistry.org/Part:BBa_B0015 BBa_B0015]
|double terminator
|2012/1/23L
|pSB1AK3
|-
|}


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Revision as of 08:16, 22 July 2012

Project name <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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Result Ligation

Plate colonies?
Ligation pur +++
Ligation 1/10 ~280
positive transformation control (pSB1A2::R0011) +++
negative ligation control (pSC002 dig. ie. insert) 0
negative ligation control (pSC003 dig. ie. vector) ~240
negative transformation control (NEB only) 0

There is a background of self ligation, I expect that 1 over 10 bacteria have the wrong plasmid. So I make a PCR colony, and I make a culture of colonies I PCR.

  • Tubes L1->L12 are clones from ligation plates.
  • Tubes T-1->T-3 are clones from negative control plate (vector)

Here is the content of 1 tube of PCR :

  • VR (diluted 10 times from the stock tube) : 2,5µL
  • VF2 (diluted 10 times from the stock tube) : 2,5µL
  • PCR MM 2X : 25 µL
  • water : 20 µL

Here is the PCR pgm :

  • 95°C - 15' (to lyse cells too)
  • 95°C - 30"
  • 50°C - 30"
  • 72°C - 40"
  • 72°C - 10'
  • 4°C -[math]\displaystyle{ \infty }[/math]

-> 24 cycles

Biobricks retrieval

Parts number Name Distribution/Plate/Well Backbone
BBa_I14034 P(Kat) 2012/2/13B pSB2K3
BBa_B0034 RBS 2012/1/2M pSB1A2
BBa_B0015 double terminator 2012/1/23L pSB1AK3
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