Team:Calgary/Notebook/Protocols/Process4
From 2011.igem.org
(Difference between revisions)
Emily Hicks (Talk | contribs) |
Emily Hicks (Talk | contribs) |
||
Line 37: | Line 37: | ||
<p>The following cylcing conditions are used.</p> | <p>The following cylcing conditions are used.</p> | ||
- | + | <CENTER> <table border="1px"> | |
<tr> | <tr> | ||
Line 43: | Line 43: | ||
</tr> | </tr> | ||
- | |||
<tr> | <tr> | ||
<td><CENTER>2 min at 50°C</CENTER></td> | <td><CENTER>2 min at 50°C</CENTER></td> | ||
</tr> | </tr> | ||
+ | |||
<tr> | <tr> | ||
<td><CENTER>10 min at 95°C</CENTER></td> | <td><CENTER>10 min at 95°C</CENTER></td> | ||
</tr> | </tr> | ||
+ | |||
<tr><td><b>40 cycles of:</b></tr></td> | <tr><td><b>40 cycles of:</b></tr></td> | ||
- | <tr> | + | |
+ | <tr> | ||
<td><CENTER>15 sec at 95°C</CENTER></td> | <td><CENTER>15 sec at 95°C</CENTER></td> | ||
</tr> | </tr> | ||
- | + | ||
+ | <tr> | ||
<td><CENTER>1 min at 60°C</CENTER></td> | <td><CENTER>1 min at 60°C</CENTER></td> | ||
</tr> | </tr> | ||
- | + | ||
+ | <tr> | ||
<td><b>Melting curve analysis</b></td> | <td><b>Melting curve analysis</b></td> | ||
</tr> | </tr> | ||
- | <tr> | + | |
+ | <tr> | ||
<td><CENTER>15 sec at 95°C</CENTER></td> | <td><CENTER>15 sec at 95°C</CENTER></td> | ||
</tr> | </tr> | ||
- | </table></CENTER></html>}} | + | |
+ | </table> | ||
+ | </CENTER> | ||
+ | </html>}} |
Revision as of 21:55, 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 |