Editing
Noi/NOTES/2012-8-3
(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!
== Amplification of bisulfite sequencing libraries with N2 barcoded primers == {| {{table}} border = 1 | width="160pt" align="center" style="background:#f0f0f0;"|'''Components''' | width="60pt" align="center" style="background:#f0f0f0;"|'''1 rxn''' | align="center" style="background:#f0f0f0;"|'''26rxn mix''' |- | DNA template||10.00||0.00 |- | 10uM PCR_F||2.00||52.00 |- | 10uM PCR_R.Indx (1-8)||2.00||0.00 |- | 2x iQ Supermix||50.00||1,300.00 |- | H2O||36.00||936.00 |} * Aliquot 88ul, add 2ul of 10uM PCR_R Ind (Ind1-8), 10ul of bis-cvt DNA template '''Program''' 98C 2min (98C 15sec -> 64C 20sec -> 72C 30sec) x10cycles -> 72C 3min -> 15CHold {| {{table}} | width="80pt" align="center" style="background:#f0f0f0;"|'''Sample ID''' | width="70pt" align="center" style="background:#f0f0f0;"|'''Labeling''' | align="center" style="background:#f0f0f0;"|'''Indx''' |- | WGBS.GM12891||#1||PCR_R.N2Indx1 |- | WGBS.GM12892||#2||PCR_R.N2Indx2 |- | WGBS.GM10861||#3||PCR_R.N2Indx3 |- | WGBS.GM11994||#4||PCR_R.N2Indx4 |- | WGBS.GM11995||#5||PCR_R.N2Indx5 |- | WGBS.GM18506||#6||PCR_R.N2Indx6 |- | WGBS.GM18507||#7||PCR_R.N2Indx7 |- | WGBS.GM18505||#8||PCR_R.N2Indx8 |} '''qPCR results'''<br> [[File:20120801_qPCRWGBS1-8-1.png| 700px]] <br> * Purified sequencing libraries with 0.8x vol AMPure bead with epMotion (3 strips), eluted with 25ul EB for each well -> total volume at the end ~70ul '''Report file of epMotion run:'''[[Media:MasterReport_20120802_beadPure.txt]]<br> [[File:BeadPure_Aug2.png| 500px]] [[File:ZhangLab_2 2012-08-02 16hr 05min_bead-HAPMAP-WGBS.jpg| 400px]] <br> * Note: To validate the quality of sequencing library, Dr. Zhang suggested to do small scale sequencing before doing deep sequencing, so I pooled small amount of each sequencing library and performed PAGE-size selection.
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