Editing
Dinh:TAB Protocol
(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!
== TAB-Seq protocol == * This protocol cannot start from methylated adaptor ligated libraries. The 5mC would be converted to T. * Protocol guides: Nature protocol for Tet-assisted bisulfite sequencing of 5hmC : http://www.nature.com/nprot/journal/v7/n12/abs/nprot.2012.137.html Manual for kits * T4 Phage Ξ²-glucosyltransferase (T4-BGT) from NEB ($65.63) T4 Phage Ξ²-glucosyltransferase NEB buffer 4 (10x) Uridine Diphosphate Glucose (50X) * Tet1 Oxidation kit from Wisegene ($669 + $90 shipping) Tet1 oxidation reagent 1 Tet1 oxidation reagent 2 Tet1 protein Proteinase K * Misc: Micro Bio-Spin 30 columns (BioRad) - By gravitational force, larger fragments will get pulled down first, smaller fragments are retained in the gel. - Use P6 columns may be better because it have a smaller pore size, so cleaner separation can be achieved. EpiTect Bisulfite Kit PfuTurbo Cx Hotstart DNA polymerase (agilent) CpG Methyltranferase (M.SssI, NEB), $65 QIAEX II gel extraction kit (150), $172 - These columns are cheaper than QiaQuick and the MinElute columns, but seems to have better flexibility in purifying larger fragments - Since I do not want to perform Covaris shearing on the DNA for BSPP, I should use the QIAEX II columns instead of the QiaQuick/MinElute columns.
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