Team:Calgary/Notebook/Protocols/Process4
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<p>We have a mastermix of PCR enzyme, sybr green, and dNTPs that is 2x. Primers will be stored at 100μM and used at final concentrations between 200-600nM. The PCR mix will be as follows:</p> | <p>We have a mastermix of PCR enzyme, sybr green, and dNTPs that is 2x. Primers will be stored at 100μM and used at final concentrations between 200-600nM. The PCR mix will be as follows:</p> | ||
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- | <td | + | <td><div class="heading"><p style="font-size:14px; font-weight:bold"><a style="text-decoration:none" name="qRT-PCR"></a> qRT-PCR using Quanta PCR mastermix </p></div></td> |
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<td><b>Component</b></td> | <td><b>Component</b></td> | ||
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<p>The following cylcing conditions are used.</p> | <p>The following cylcing conditions are used.</p> | ||
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- | <td | + | <td><p style="font-size:14px; font-weight:bold"><a style="text-decoration:none" name="reaction conditions"></a> </p></div></td> |
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<td><CENTER>2 min at 50°C</CENTER></td> | <td><CENTER>2 min at 50°C</CENTER></td> |
Revision as of 21:54, 27 September 2011
qRT-PCR
We have a mastermix of PCR enzyme, sybr green, and dNTPs that is 2x. Primers will be stored at 100μM and used at final concentrations between 200-600nM. The PCR mix will be as follows:
Component | Volume |
Quanta PCR master mix | 5 uL |
Primers (Fwd&Rev) at 600nM each | 3 uL |
cDNA | 2 uL |
Final Volume | 10 uL |
The following cylcing conditions are used.
40 cycles of: |
Melting curve analysis |