Team:Tuebingen/Notebook/Protocols/pgem-ligation
From 2013.igem.org
(Difference between revisions)
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- | <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: | + | <a href="https://2013.igem.org/Team:Tuebingen/Notebook/Protocols" style="font-size: 18px">Back to Protocols</a> |
+ | <p> </p> | ||
+ | |||
+ | <h3>Reagents</h3> | ||
+ | <table border="0"> | ||
+ | <colgroup> | ||
+ | <col width="80"> | ||
+ | <col width="300"> | ||
+ | </colgroup> | ||
+ | |||
+ | <tr> | ||
+ | <td style="text-align: center">5 µL</td> | ||
+ | <td>2x Rapid Ligation Buffer</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td style="text-align: center">0.5 µL (25 ng)</td> | ||
+ | <td>pGEM vector</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td style="text-align: center">3.5 µL</td> | ||
+ | <td>PCR</td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td style="text-align: center">10 mL</td> | ||
+ | <td>KCl (c = 250 mM)</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | <p> </p> | ||
+ | |||
+ | <h3>Procedure</h3> | ||
+ | <p>Dissolve all reagents in 1000 mL Aqua dest., adjust pH to 7.0 (with NaOH). Sterilize by autoclaving. After autoclaving add 5 mL MgCL2 (c = 2 M).</p> | ||
</div> | </div> |
Revision as of 02:44, 4 October 2013
pGEM Ligation
Back to Protocols
Reagents
5 µL | 2x Rapid Ligation Buffer |
0.5 µL (25 ng) | pGEM vector |
3.5 µL | PCR |
10 mL | KCl (c = 250 mM) |
Procedure
Dissolve all reagents in 1000 mL Aqua dest., adjust pH to 7.0 (with NaOH). Sterilize by autoclaving. After autoclaving add 5 mL MgCL2 (c = 2 M).