Editing
Matt:LabNotes/2014-5-22
(section)
Jump to navigation
Jump to search
Warning:
You are not logged in. Your IP address will be publicly visible if you make any edits. If you
log in
or
create an account
, your edits will be attributed to your username, along with other benefits.
Anti-spam check. Do
not
fill this in!
===Lambda Exonuclease Digestion=== *Divide each tube of 220ul into two 1.5ml tubes of 110ul with total amplicon of ~30ug {| {{table}} | align="center" style="background:#f0f0f0;"|'''Components''' | align="center" style="background:#f0f0f0;"|'''Volume (ul)''' |- | Amplicon||110 |- | 10X Lambda Exo Buffer||15 |- | Lambda Exonuclease||10 |- | H2O||15 |- | total||150 |- | |} *Incubated at 37C for 1hr *Purified each of the tubes with 3 columns (6 columns total) of Zymo ssDNA/RNA kit *Eluted with 27ul each *Recombined and measured ssDNA with Nanodrop: **49.2ng/ul x 160ul = 7.87ug (12% yield) ***Why is it so low? Expected to be about 40%... **Low yield could be because Zymo kit is for ssDNA between 17nt-200nt and these oligos are slightly above 200nt before USER & DpnII digestion *Noi suggests I may have had too much DNA in each tube for lambda digestion. She says 10ug DNA for 100ul reaction (15ug/150ul reaction) <!-- To continue see: [[Matt:LabNotes/2013-5-13]] --> *Continued on: [[Matt:LabNotes/2014-6-24]]
Summary:
Please note that all contributions to ZhangLabWiki may be edited, altered, or removed by other contributors. If you do not want your writing to be edited mercilessly, then do not submit it here.
You are also promising us that you wrote this yourself, or copied it from a public domain or similar free resource (see
ZhangLabWiki:Copyrights
for details).
Do not submit copyrighted work without permission!
Cancel
Editing help
(opens in new window)
Navigation menu
Personal tools
Not logged in
Talk
Contributions
Create account
Log in
Namespaces
Page
Discussion
English
Views
Read
Edit
View history
More
Search
Navigation
Main Page
Current events
Recent changes
Random page
Investigators
Matt Cai
Song Chen
Eric Chu
Dinh Diep
Elizabeth Duong
Shicheng Guo
Alan Fung
Daniel Jacobsen
Blue Lake
Huy Lam
Alice Li
Andrew Richards
Brandon Sos
Chris Wei
Yan Wu
Kun Zhang
Tools
What links here
Related changes
Special pages
Page information