Team:Exeter/lab book/proto/7

From 2012.igem.org

(Difference between revisions)
(Created page with "{{Template:Team:Exeter/e-candi_banner}} <html xmlns="http://www.w3.org/1999/xhtml"> <head> <meta http-equiv="Content-Type" content="text/html; charset=utf-8" /> <title>Protocol ...")
Line 68: Line 68:
       <li><p>31µL MilliQ H2O</p></li>
       <li><p>31µL MilliQ H2O</p></li>
     </ul>
     </ul>
 +
   
 +
    <br>
 +
   
     <p>Run a two-step PCR experiment using the following settings:</p>
     <p>Run a two-step PCR experiment using the following settings:</p>
     <ul type="square">
     <ul type="square">

Revision as of 12:36, 19 September 2012

Protocol 7

Two-Step PCR

Make-up a 50µL total solution in a PCR tube of:

  • 5µL of 10ng/µL genomic DNA template

  • 1µL dNTPs

  • 1µL Forward Primer (@100pmol/µL)

  • 1µL Reverse Primer (@100pmol/µL)

  • 10µL High-Fidelity Phusion 5x Buffer

  • 1µL High-Fidelity Phusion polymerase

  • 31µL MilliQ H2O


Run a two-step PCR experiment using the following settings:

  • Stage 1: 98oC for 30 seconds for 1 cycle

  • Stage 2: 98oC for 10 seconds followed by 72oC for 20 seconds for 30 cycles

  • Stage 3: 72oC for 5 minutes for 1 cycle

Retrieved from "http://2012.igem.org/Team:Exeter/lab_book/proto/7"