PrbbBB:colony pcr v1: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Line 23: | Line 23: | ||
<table frame=box> | <table frame=box> | ||
<tr align=right> | <tr align=right> | ||
<td></td> <th width=100>11µl single reaction</th> <th width=100>60xMaster</th> <th width=100> | <td></td> <th width=100>11µl single reaction</th> <th width=100>60xMaster</th> <th width=100>115xMaster</th></tr> | ||
<tr align=right> <td>H2O</td> <td>7.5µl</td> <td></td> <td></td> </tr> | <tr align=right> <td>H2O</td> <td>7.5µl</td> <td>450</td> <td>862</td> </tr> | ||
<tr align=right> <td>10x Amplitaq buffer</td> <td>1.1µl</td> <td></td> | <tr align=right> <td>10x Amplitaq buffer</td> <td>1.1µl</td> <td>66</td> <td>126.5</td> </tr> | ||
<tr align=right> <td>10mM dNTP</td> <td>0.22µl</td> <td></td> | <tr align=right> <td>10mM dNTP</td> <td>0.22µl</td> <td>13.2</td> <td>25.3</td> </tr> | ||
<tr><td></td></tr> | <tr><td></td></tr> | ||
<tr align=right> <td>VF2 primer 10 µM</td> <td>0.55µl</td> <td></td> | <tr align=right> <td>VF2 primer 10 µM</td> <td>0.55µl</td> <td>33</td> <td>63.3</td> </tr> | ||
<tr align=right> <td>VR primer 10 µM</td> <td>0.55µl</td> <td></td> | <tr align=right> <td>VR primer 10 µM</td> <td>0.55µl</td> <td>33</td> <td>63/3</td> </tr> | ||
<tr align=right> <td>AmpliTaq 5U/µl</td> <td>0.1µl</td> <td></td> | <tr align=right> <td>AmpliTaq 5U/µl</td> <td>0.1µl</td> <td>6</td> <td>11.5</td> </tr> | ||
<tr><td></td></tr> | <tr><td></td></tr> | ||
<tr align=right> <td>template DNA</td> <td>1µl</td> <td>--</td> | <tr align=right> <td>template DNA</td> <td>1µl</td> <td>--</td> <td>--</td> </tr> | ||
</table> | </table> | ||
</td> | </td> |
Revision as of 04:58, 26 February 2009
Overview
Materials
- 96-well PCR plate
- AmpliTaq DNA Polymerase 5 U/µl (Roche)
- AmpliTaq Buffer 10 x
- dNTP mix 10mM each nucleotide
- ddH2O
- primers:
Procedure
I PCR reaction
|
| ||||||||||||||||||||||||||||||||||||||||||||||
II Agarose Gel
Notes
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
raik: no comment
References
Contact
or instead, discuss this protocol.