User:University of Washington/8 September 2008

From 2008.igem.org

(Difference between revisions)
 
Line 1: Line 1:
 +
==LuxR from AraC and TetR(Faifan)==
 +
 +
-Sequences of LuxR construct came back. 4/8 contained P1010, the other 4/8 had too many N's-cannot determine.
 +
 +
-Screening 14 colonies by colony PCR
 +
*resuspended each colony into 50 ul dH2O, vortexed
 +
*mix (10 ul rxn)
 +
** 5.95 ul dH2O
 +
** 1 ul 10x Taq Buffer
 +
** 0.25 ul 10mM dNTPs
 +
** 0.50 ul MgCl2
 +
** 0.6 ul VF2(10 pmol/ul)
 +
** 0.6 ul VR(10 pmol/ul)
 +
** 0.1 ul Taq
 +
** 1 ul colony suspension
 +
*Thermocycling
 +
**1 cycle : 95 C : 6 mins
 +
**35 cycle: 95 C : 30 s
 +
**________: 53 C : 30 s
 +
**________: 72 C : 2 mins
 +
**1 cycle : 72 C : 10 mins
 +
**1 cycle : 4 C  : forever
 +
----
----
Back to [[Team:University_of_Washington/Notebook#Notebook]]
Back to [[Team:University_of_Washington/Notebook#Notebook]]

Latest revision as of 08:13, 9 September 2008

LuxR from AraC and TetR(Faifan)

-Sequences of LuxR construct came back. 4/8 contained P1010, the other 4/8 had too many N's-cannot determine.

-Screening 14 colonies by colony PCR

  • resuspended each colony into 50 ul dH2O, vortexed
  • mix (10 ul rxn)
    • 5.95 ul dH2O
    • 1 ul 10x Taq Buffer
    • 0.25 ul 10mM dNTPs
    • 0.50 ul MgCl2
    • 0.6 ul VF2(10 pmol/ul)
    • 0.6 ul VR(10 pmol/ul)
    • 0.1 ul Taq
    • 1 ul colony suspension
  • Thermocycling
    • 1 cycle : 95 C : 6 mins
    • 35 cycle: 95 C : 30 s
    • ________: 53 C : 30 s
    • ________: 72 C : 2 mins
    • 1 cycle : 72 C : 10 mins
    • 1 cycle : 4 C  : forever


Back to Team:University_of_Washington/Notebook#Notebook

Retrieved from "http://2008.igem.org/User:University_of_Washington/8_September_2008"