User:Jose Fabricio Lopez/Notebook/Logbook/2011/06/09: Difference between revisions

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**Lane 3.3μL dye & 5μL Control (Tube C from Daniel´s extraction).
**Lane 3.3μL dye & 5μL Control (Tube C from Daniel´s extraction).


[[Image:Gel.07junio11.Fibo.PCR.pSB1t3purificacion.JPG|thumb|left]]
[[Image:Gel.07junio11.Fibo.PCR.pSB1t3purificacion.JPG|thumb|center]]
 
*Ligation of PCR purification:
 
I made two ligations by distinct ways.
 
**DNA insert  5μL
**DNA vector  1μL
**Buffer10X  1μL
**H<sub>2</sub>O        1μL
**T4LigaseNEB 1μL
 
Second one
**DNA insert  2μL
**DNA vector  3μL
**Buffer10X  1μL
**H<sub>2</sub>O        3μL
**T4LigaseNEB 1μL
 




Revision as of 12:35, 10 June 2011

iGEM 2011 <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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Objective

  • Clean and know purity of Daniel´s PCR

WetLab

  • Purification of PCR product with Roche´s kit of High Pure PCR product.
    • I joined tube A and B and were filled to 100μL
  • See purity of PCR cleaning with a gel.
    • Lane 1.5μL Ladder
    • Lane 2.3μL dye & 5μL DNA of cleaned PCR.
    • Lane 3.3μL dye & 5μL Control (Tube C from Daniel´s extraction).
  • Ligation of PCR purification:

I made two ligations by distinct ways.

    • DNA insert 5μL
    • DNA vector 1μL
    • Buffer10X 1μL
    • H2O 1μL
    • T4LigaseNEB 1μL

Second one

    • DNA insert 2μL
    • DNA vector 3μL
    • Buffer10X 1μL
    • H2O 3μL
    • T4LigaseNEB 1μL
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