Team:University of Ottawa/14 July 2008

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(New page: __TOC__ ==Today in the Lab== '''Matt''' :'''PCR Amplification''' ::<li> Corey decided it would be best to start from the beginning and amplify the PTP2 out of Genomic DNA again. I redid th...)
 
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__TOC__
__TOC__
==Today in the Lab==
==Today in the Lab==
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::<li> 97 plasmid was inoculated from master plate so that it can glycerol stocked for tomorrow.
::<li> 97 plasmid was inoculated from master plate so that it can glycerol stocked for tomorrow.
:'''PCR Confirmation'''
:'''PCR Confirmation'''
-
::<li> Corey wanted me to do another PCR confirmation of the 97 plasmid with primers F58,F59. Expecting a band size of 2200 for this PCR.
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::<li> Corey wanted me to do another PCR confirmation of the 97 plasmid with primers F58,F59. Expecting a band size of 2200 for this PCR when I run it on a gel for tommorrow.
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'''Chris'''
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:'''Digestion of AtCRE (again)'''
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::<li> AtCRE was digested with EcoRI again for one hour at 37 C
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::<li> the result was run on a 1% gel, resulting in five bands where only two were expected.
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::<li> using Vector NTI, we figured out that the incorrect enzyme was used for the digestion; in fact, EagI should have been used instead.
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'''Dan'''
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:'''PCR amplification of construct 1'''
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::<li> The amplification product was run on a gel however no clear bands showed up on the kodac machine. However the correct band was seen on the cutting UV machine.
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::<li> The band was extracted using the Sigma kit and eluted with water.
 +
::<li> Absorbance measurements indicated that no DNA was obtained, next time elution solution at 65 C will be used
 +
::<li> PCR amplification of 0A and 0B was run overnight.

Latest revision as of 18:31, 30 July 2008

Untitled Document

 

 


Contents

Today in the Lab

Matt

PCR Amplification
  • Corey decided it would be best to start from the beginning and amplify the PTP2 out of Genomic DNA again. I redid the PCR and I am running it overnight.
    • Inoculation
  • 97 plasmid was inoculated from master plate so that it can glycerol stocked for tomorrow.
    • PCR Confirmation
  • Corey wanted me to do another PCR confirmation of the 97 plasmid with primers F58,F59. Expecting a band size of 2200 for this PCR when I run it on a gel for tommorrow.

    • Chris

    Digestion of AtCRE (again)
  • AtCRE was digested with EcoRI again for one hour at 37 C
  • the result was run on a 1% gel, resulting in five bands where only two were expected.
  • using Vector NTI, we figured out that the incorrect enzyme was used for the digestion; in fact, EagI should have been used instead.

    • Dan

    PCR amplification of construct 1
  • The amplification product was run on a gel however no clear bands showed up on the kodac machine. However the correct band was seen on the cutting UV machine.
  • The band was extracted using the Sigma kit and eluted with water.
  • Absorbance measurements indicated that no DNA was obtained, next time elution solution at 65 C will be used
  • PCR amplification of 0A and 0B was run overnight.
    • Retrieved from "http://2008.igem.org/Team:University_of_Ottawa/14_July_2008"