Editing
Jie:LabNotes/CpgSeq/2009-5-29
(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!
==PCR with AmpF/R6.2Sol and iProof== x10 Template 5ul 2X iProof Mastermix 50ul 500ul AmpF6.2SoL (10uM) 4ul 40ul AmpR6.2SoL (10uM) 4ul 40ul 50X SYBG I 0.8ul 8ul H2O 36.2ul 362ul 98C 30S -> (98C 10S -> 58C 20S -> 72C 20S) x 8 -> (98C 10S -> 72C 20S) x 10-> 72C 5 min -> 15C hold. I purified the amplicons with Qiaquick. Gel purified the cpg30k four subsets amplicons and normalized the final amplicons with 1:1 ratio. The 09_8 and 09_9 cpg97k amplicons concentrations are: 09_8: 11.3ng/ul x 30ul; 09_9: 13.7ng/ul x 30ul. ==PCR amplification with AmpF6.3NH2/AmpR6.3NH2 and dUTP:dNTP 1:40 I did the dUTP_PCR with template from the mixture captured targets, the Qiaquick purified captured targets of 97k, and the directed captured targets of 97k. reaction system x8 x6 H2O 42.6ul 340.8ul 2x Master mix 50ul 400ul dUTP(1mM) 2ul 16ul AmpF6.3NH2(10uM) 2ul 16ul AmpR6.3NH2(10uM) 2ul 16ul 50x SYBG I 0.4ul 3.2ul template 0.25ul/each for cpg30k mixture, 0.05ul/each for purified cpg97k, 1ul/each for direct cpg97k. Total 100ul 1400ul 94C 3min -> 9cycles of (94C 45sec -> 55C 45sec -> 72C 45sec) -> 72C 3min -> 4C Qiaquick purification: No_09_8_30k: 34ng/ul x 60ul; No_09_8_97k: 21ng/ul x 60ul; No_09_9_30k: 36ng/ul x 60ul; No_09_8_97k: 24ng/ul x 60ul;
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