Team:University of Ottawa/30 July 2008

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(New page: ='''Today in the Lab'''= '''Tammy''' <TABLE BORDER=2> <TR> <TD>'''Sample'''</TD><TD>'''Concentration (ng/μL)'''</TD><TD>'''V (μL)'''</TD><TD>'''Total DNA (ng)'''</TD> </TR> <TR> <TD...)
 
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='''Today in the Lab'''=
='''Today in the Lab'''=
'''Tammy'''
'''Tammy'''
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<TR> <TD>'''Total'''</TD><TD>'''30'''</TD> </TR>
<TR> <TD>'''Total'''</TD><TD>'''30'''</TD> </TR>
</TABLE>
</TABLE>
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=='''Chris'''==
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'''PCR Confirmation'''
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:<li> Ran all 5 PCR product samples on a 1% gel for 40 minutes at 80 V
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:<li> Desired band appeared on gel; PCR amplification was successful.
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:<li> Used PCR cleanup kit to purify samples
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:<li> Measured absorbance of PTP2 samples
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'''Digestion of PTP2'''
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:<li> Digested PTP2 with BamHI and XhoI; ran 5 tubes in parallel. Incubated at 37 C for on hour. Ran one water control.
 +
:<li> Used PCR cleanup kit to purify digestion products. Measured absorbance to determine concentration.
 +
:<li> Concentrations too low for ligation.
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=='''Matt'''==
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:<li> Measured absorbance of pSSA42 gel extraction products from digestion.
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=='''Dan'''==
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'''Gel Extraction'''
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:<li>T123 amplification product was run on a gel and extracted.
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:<li>24 gel extraction were performed in total, care was taken to minimize UV exposure (samples were not exposed for longer than 20s)
 +
'''Gel of extracted PCR amplification product'''
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:<li> Expected bands were seen, the T123 is good to go.
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'''0A and 0B absorbances'''
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:<li> absorbances of 0A and 0B indicated very good concentrations
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'''Ligation and digestion'''
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:<li> Digestion (with SphI and PstI) of T123 0A and 0B were all performed and left overnight

Latest revision as of 14:57, 12 August 2008

Untitled Document

 

 


Contents

Today in the Lab

Tammy

SampleConcentration (ng/μL)V (μL)Total DNA (ng)
pSSA141297903
OB17061020


  • Digestion Step 1 of pSSA14 with PstI
    • Digestion Reaction componentsVol (μl)
    H2O4
    Buffer 31.5
    BSA1.5
    PstI1
    DNA template7
    Total15


  • Digestion Step 1 of OB with PstI
    • Digestion Reaction componentsVol (μl)
    H2O5
    Buffer 31.5
    BSA1.5
    PstI1
    DNA template6
    Total15


  • Digestion Step 2 of pSSA14 with ClaI
    • Digestion Reaction componentsVol (μl)
    H2O9.5
    Buffer 43
    BSA1.5
    ClaI1
    DNA template15
    Total30


  • Digestion Step 2 of OB with ClaI
    • Digestion Reaction componentsVol (μl)
    H2O9.5
    Buffer 43
    BSA1.5
    ClaI1
    DNA template15
    Total30

    Chris

    PCR Confirmation

  • Ran all 5 PCR product samples on a 1% gel for 40 minutes at 80 V
  • Desired band appeared on gel; PCR amplification was successful.
  • Used PCR cleanup kit to purify samples
  • Measured absorbance of PTP2 samples

    • Digestion of PTP2

  • Digested PTP2 with BamHI and XhoI; ran 5 tubes in parallel. Incubated at 37 C for on hour. Ran one water control.
  • Used PCR cleanup kit to purify digestion products. Measured absorbance to determine concentration.
  • Concentrations too low for ligation.

    • Matt

  • Measured absorbance of pSSA42 gel extraction products from digestion.

    • Dan

    Gel Extraction

  • T123 amplification product was run on a gel and extracted.
  • 24 gel extraction were performed in total, care was taken to minimize UV exposure (samples were not exposed for longer than 20s)

    • Gel of extracted PCR amplification product

  • Expected bands were seen, the T123 is good to go.

    • 0A and 0B absorbances

  • absorbances of 0A and 0B indicated very good concentrations

    • Ligation and digestion

  • Digestion (with SphI and PstI) of T123 0A and 0B were all performed and left overnight
    • Retrieved from "http://2008.igem.org/Team:University_of_Ottawa/30_July_2008"