Editing
Blue:RNA-Seq Experiments:11062013
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!
== Purpose == *Use bioanalyzer to test different fragmentation/poly(A) conditions - continuation on 11052013 == RNAseIII-Fragmented Library Preparation == :'''Conditions''' #RNaseIII (5mM MnCl2)/PAP (2.5mM MnCl2 and 1mM ATP) 10min #RNaseIII (5mM MgCl2)/PAP (2.5mM MnCl2 and 0.1mM ATP) 10min #RNaseIII (5mM MnCl2)/PAP (2.5mM MnCl2 and 1:1 dATP) 10min #RNaseIII (5mM MnCl2)/PAP (2.5mM MnCl2 and 1:1 dATP) 30min #RNaseIII (10mM MnCl2)/PAP (5mM MnCl2 and 1:1 dATP) 10min #RNaseIII (10mM MnCl2)/PAP (5mM MnCl2 and 1:1 dATP) 30min #RNaseIII (20mM MnCl2)/PAP (10mM MnCl2 and 1:1 dATP) 10min #RNaseIII (20mM MnCl2)/PAP (10mM MnCl2 and 1:1 dATP) 30min :'''RNA''' ::{| {{table}} |- |||||||||||<u>Volume</u> |- |UHRR 10ng/ul ||||||||||1ul |- |5x Ambion PAP buffer||||||||||0.08ul |- |MnCl2||||||||||0.4ul |- |RNase III||||||||||0.1ul |- |dH20||||||||||0.42ul |- |Total||||||||||2ul |- |} :::''Incubate @ 37C 5 min'' :'''3' Tailing''' ::{| {{table}} |- |||||||||||<u>Volume</u> |- |fRNA||||||||||2ul |- |5x Ambion PAP Buffer||||||||||0.72ul |- |ATP (or mix)||||||||||0.25ul |- |2U/ul PAP||||||||||0.2ul |- |40U/ul Rnase Inhibitor||||||||||0.2ul |- |dH2O||||||||||0.63ul |- |} :::''Incubate @ 37C 10 min - 30 min'' :::''Column Purify Samples'' Bioanalyzer: [[File:2100_expert_Eukaryote_Total_RNA_Pico_DE72903057_2013-11-08_08-37-03.pdf]]
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)
Template used on this page:
Template:Table
(
edit
)
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