Team:Macquarie Australia/Notebook3
From 2010.igem.org
(Difference between revisions)
Line 86: | Line 86: | ||
+ | <li type="disc"> The pET-3A vector plasmid that is to be used for the transformation needs to be purified </li> | ||
<li type="disc"> | <li type="disc"> | ||
- | + | The pET-3A plasmid was purified using the Qiagen Plasmid Midi Prep Kit as per the manufacturer’s protocols</li> | |
- | + | <li type="disc"> | |
- | + | The only difference made to the protocol is that only 5ml of Buffer QC was used on the 2nd wash step</li> | |
+ | <li type="disc"> | ||
+ | The plasmid (two samples A & B) as well as some column flow-through collected at different stages of the purification were run on a GelRed post-stained 1.5% agarose gel and photo taken for visualization (see below).</li> | ||
+ | <li type="disc"> | ||
+ | A NanoDrop spectrophotometer reading was also recorded to check the quality of the extracted genomic DNA.</li> | ||
+ | <li type="disc"> | ||
+ | There was no DNA pellet formed at the end of this protocol so we may have lost some DNA</li> | ||
+ | <li type="disc"> | ||
+ | There was also some plasmid observed in some of the column flow through so some plasmid has been lost</li> | ||
Revision as of 09:26, 24 October 2010
PROJECT LAB BOOK
Welcome to the Macquarie University project lab book page!
DAY-BY-DAY PROGRESS FOR THE TRANSFORMATION AND CLONING
22nd September 2010
Plasmid Prep using Qiagen Plasmid Midi Prep Kit