Editing
Daniel:Notebook/RNAFACS/2016-1-12
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!
=Poly T Primer Test ([[Daniel:Notebook/RNAFACS/2016-1-11|Started 01/11/16]])= [[Daniel:Notebook/RNAFACS|Back to Calendar]] ==Buffers== *Wash Buffer A **200 uL Stellaris RNA FISH Wash Buffer A (Biosearch Technologies Cat# SMF-WA1-60) **Add 700 uL Nuclease-free water **Add 100 uL Deionized Formamide **Mix well by vortexing gently *Wash Buffer A w/DAPI **200 uL Stellaris RNA FISH Wash Buffer A (Biosearch Technologies Cat# SMF-WA1-60) **Add 700 uL Nuclease-free water **Add 100 uL Deionized Formamide **Add 1 uL of 20 ug/mL DAPI stock (dilute original 1:1000 and store in 4C) **Mix well by vortexing gently ==Protocol== #Aspirate out hybridization buffer and add 1 mL of Wash Buffer A #Incubate in the dark at 37 °C for 30 minutes #Aspirate the Wash Buffer A, and then add 1 mL of DAPI nuclear stain (Wash Buffer A consisting of 5 ng/mL DAPI) to counterstain the nuclei #Incubate in the dark at 37 °C for 30 minutes #Aspirate the DAPI staining buffer, and then add 1 mL of Wash Buffer B. Incubate at room temperature for 2-5 minutes #Add a small drop (approximately 15 μL) of Vectashield Mounting Medium onto center of dish #Seal the coverglass perimeter with clear nail polish, and allow to dry #If necessary, gently wipe away any dried salt off the coverglass with water
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