Team:University of Ottawa/29 July 2008

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=='''Tammy and Dan'''==
=='''Tammy and Dan'''==
:'''0.8% Agarose Gel Electrophoresis of T123 PCR Products'''
:'''0.8% Agarose Gel Electrophoresis of T123 PCR Products'''
-
::<li> Each PCR reaction tube (50 &mu;L) was divided into 2 and ran on 2 separate wells
+
::<li> Each PCR reaction tube (50 &mu;L) was divided into 2 and ran on 2 separate wells to decrease DNA load and to ensure identification of appropriate band size
:'''Agarose Gel Extraction'''
:'''Agarose Gel Extraction'''
 +
::<li> 24 bands were cut from the gel using minimal UV exposure
 +
::<li> 6 gel bands were pooled into 1 column for a total of 4 aliquots of purified T123 DNA
 +
 +
=='''Matt'''==
 +
:'''Inoculation'''
 +
::<li> Inoculation of pSSA42 turned out nicely, control was clean - performed a miniprep from inoculation.
 +
:'''Transformation'''
 +
::<li> Plates of transformation in competent cells did not yield any colonies - control was clean.
 +
:'''Digestion'''
 +
::<li> Digestion of pSSA42 was performed with BamHI + xhoI.

Latest revision as of 18:52, 7 August 2008

Untitled Document

 

 

Today in the lab

Contents

Chris

PCR Amplification of PTP2
  • Began running experimentation alongside Matt to increase chances of success
  • Ran 5 samples, including two water controls as per Cory's request.
  • Master mix: 50 uL Buffer, 5 uL dNTP, 12.5 uL F60 and F61, 2.5 uL DNA (at 25 ng/uL), 142.5 uL water.
    • Digestion of PSSA42
  • Ran five samples and one water control
  • Per tube: 3 uL water, 3 uL Buffer 3, 3 uL BSA, 1.5 uL XhoI and BamHI, 16 uL DNA template.
  • Incubated at 37 C for one hour
    • PCR Cleanup
  • Used PCR cleanup kit to purify PTP2
  • Measured absorbance of resulting DNA samples. The concentrations were found to be very low; PCR did not work. It was later determined that DNA template was not added, by accident.
    • PCR Amplification of PTP2
  • Ran PCR with same constraints as previously. Let run overnight.

    • Tammy and Dan

    0.8% Agarose Gel Electrophoresis of T123 PCR Products
  • Each PCR reaction tube (50 μL) was divided into 2 and ran on 2 separate wells to decrease DNA load and to ensure identification of appropriate band size
    • Agarose Gel Extraction
  • 24 bands were cut from the gel using minimal UV exposure
  • 6 gel bands were pooled into 1 column for a total of 4 aliquots of purified T123 DNA

    • Matt

    Inoculation
  • Inoculation of pSSA42 turned out nicely, control was clean - performed a miniprep from inoculation.
    • Transformation
  • Plates of transformation in competent cells did not yield any colonies - control was clean.
    • Digestion
  • Digestion of pSSA42 was performed with BamHI + xhoI.
    • Retrieved from "http://2008.igem.org/Team:University_of_Ottawa/29_July_2008"