"
Team:Calgary/17 June 2009
From 2009.igem.org
(Difference between revisions)
Babydimples (Talk | contribs) (New page: {{Template:CalgaryHeader}} <html> <body> <style> p{ padding: 5px 5px 5px 5px; } .heading{ font: century gothic; color: #ffffff; } .name{ font: century gothic; color: #ffffff; padding: ...) |
Babydimples (Talk | contribs) |
||
| Line 137: | Line 137: | ||
<br> | <br> | ||
<div class="heading"> | <div class="heading"> | ||
| - | + | JUNE 17, 2009 | |
</div> | </div> | ||
<br> | <br> | ||
| Line 155: | Line 155: | ||
CAROL | CAROL | ||
</div> | </div> | ||
| - | |||
<div class="heading"> | <div class="heading"> | ||
| - | + | QuikChange XL Site-Directed Mutagenesis for ''luxCDABE''in TOPO vector | |
</div> | </div> | ||
<br> | <br> | ||
<div class="desc"> | <div class="desc"> | ||
</html> | </html> | ||
| - | + | Purpose: To mutate the XbaI site that is located about ~1000bp into the sequence. The reason for mutating the XbaI site for ''luxCDABE'' in TOPO vector is because to clone the sequence into a biobrick vector (pSB1AK3 and pSB1AC3), the enzyme, 'XbaI' is required for cloning purposes. Instead of cutting the sequence into two parts, we have to mutate the site so that it only cuts at the end of the sequence properly. | |
| + | * QuikChange XL Site-Directed Mutagenesis Kit retrived from Stratagene | ||
| + | * Both the control reaction and the control transformation was done as well to ensure efficiency of mutagenesis. Please see detailed procedures for mutagenesis in the protocol section. | ||
| + | * Plamids were transformed into XL Gold Ultracompetent cells (part of the kit) and was plated on LB plates overnight at 37<sup>o</sup>C. The XL Gold Ultracompetent cells allow the uptake of single stranded DNA. | ||
<html> | <html> | ||
</div> | </div> | ||
Revision as of 20:34, 29 July 2009
UNIVERSITY OF CALGARY
