Team:University of Ottawa/28 July 2008

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(New page: <TABLE BORDER=2> <TR> <TD>'''Sample'''</TD><TD>'''Concentration 1 (ng/μL)'''</TD><TD>'''V1 (μL)'''</TD><TD>'''Concentration 2 (ng/μL)'''</TD><TD>'''V2 (μL)'''</TD><TD>'''H2O (&...)
 
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='''Today in the Lab'''=
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'''Tammy and Dan'''
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<TABLE BORDER=2>
<TABLE BORDER=2>
<TR> <TD>'''Sample'''</TD><TD>'''Concentration 1 (ng/&mu;L)'''</TD><TD>'''V1 (&mu;L)'''</TD><TD>'''Concentration 2 (ng/&mu;L)'''</TD><TD>'''V2 (&mu;L)'''</TD><TD>'''H2O (&mu;L)'''</TD></TR>
<TR> <TD>'''Sample'''</TD><TD>'''Concentration 1 (ng/&mu;L)'''</TD><TD>'''V1 (&mu;L)'''</TD><TD>'''Concentration 2 (ng/&mu;L)'''</TD><TD>'''V2 (&mu;L)'''</TD><TD>'''H2O (&mu;L)'''</TD></TR>
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:<li> '''Digestion Step 1 of pSSA14 with PstI'''
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:<li> '''PCR Amplification of T123'''
<TABLE BORDER=2>
<TABLE BORDER=2>
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<TR> <TD>'''Digestion Reaction components'''</TD><TD>'''Vol (&mu;l)'''</TD> </TR>
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<TR> <TD>'''PCR Reaction components'''</TD><TD>'''1X Vol (&mu;l)'''</TD><TD>'''13X Vol (&mu;L)'''</TD> </TR>
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<TR> <TD>H2O</TD><TD>4</TD> </TR>
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<TR> <TD>5X Reaction Buffer</TD><TD>10</TD> <TD>130</TD></TR>
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<TR> <TD>Buffer 3</TD><TD>1.5</TD> </TR>
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<TR> <TD>10 mM each dNTP</TD><TD>1</TD><TD>13</TD> </TR>
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<TR> <TD>BSA</TD><TD>1.5</TD> </TR>
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<TR> <TD>F Primer (10 pmol/&mu;L) F69</TD><TD>2.5</TD><TD>32.5</TD> </TR>
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<TR> <TD>PstI</TD><TD>1</TD> </TR>
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<TR> <TD>R Primer (10 pmol/&mu;L) F70</TD><TD>2.5</TD><TD>32.5</TD> </TR>
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<TR> <TD>DNA template</TD><TD>7</TD> </TR>
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<TR> <TD>DNA Template</TD><TD>4</TD><TD>52</TD> </TR>
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<TR> <TD>'''Total'''</TD><TD>'''15'''</TD> </TR>
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<TR> <TD>Phusion Polymerase</TD><TD>0.5</TD><TD>6.5</TD> </TR>
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<TR> <TD>Filtered Sterile ddH2O</TD><TD>29.5</TD><TD>383.5</TD> </TR>
</TABLE>
</TABLE>
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::<li> Control for PCR Reaction was ddH2O instead of DNA template
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::<li> 12 PCR Reactions with identical DNA template
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'''Matt'''
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:'''PCR cleanup'''
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::<li> Performed a PCR cleanup of LiG ptp2/Pssa42
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:'''Transformation'''
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::<li> Transformed Ligated PTP2/pSSA42 into competent cells.
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:'''Inoculation'''
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::<li> Inoculated more pSSA42 vector for future digestion/ligations.

Latest revision as of 18:47, 7 August 2008

Untitled Document

 

 


Today in the Lab

Tammy and Dan

SampleConcentration 1 (ng/μL)V1 (μL)Concentration 2 (ng/μL)V2 (μL)H2O (μL)
T12375456056


  • PCR Amplification of T123
    • PCR Reaction components1X Vol (μl)13X Vol (μL)
    5X Reaction Buffer10 130
    10 mM each dNTP113
    F Primer (10 pmol/μL) F692.532.5
    R Primer (10 pmol/μL) F702.532.5
    DNA Template452
    Phusion Polymerase0.56.5
    Filtered Sterile ddH2O29.5383.5
  • Control for PCR Reaction was ddH2O instead of DNA template
  • 12 PCR Reactions with identical DNA template

    • Matt

    PCR cleanup
  • Performed a PCR cleanup of LiG ptp2/Pssa42
    • Transformation
  • Transformed Ligated PTP2/pSSA42 into competent cells.
    • Inoculation
  • Inoculated more pSSA42 vector for future digestion/ligations.
    • Retrieved from "http://2008.igem.org/Team:University_of_Ottawa/28_July_2008"