Editing
Hosuk:LabNotes/2014-10-14
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!
*[[Hosuk:Lab_Notes|LabNote]] ===Human Brain Tissue : fixing test=== *Try Human Brain Tissue fixed with 4% Formaldehyde at 4C for 12hr. ====Procedure==== *Fix 4% paraformaldehyde in 1x DEPC-PBS at 4C for 12 hr. *Wash 2x in DEPC-PBE for 5 minutes each *Glue and attach glass to MatTek dish *Permeabilize in 0.25% Triton X-100 in DEPC-treated H2O for 15 min *Wash 3 times in DEPC-H2O *Add 200 ul 0.1% pepsin in 0.1 N HCl for 15 min at 37C *Wash with 2 ml PBS three times to inactivate pepsin *Add RT mix (Hexamer, +a-dUTP) *DEPC-H2O 157uL *#M-MuLV Buffer 20uL *#25mM dNTP 2uL *#2mM a-dUTP 4uL *#100uM random hexamer RT Primer 5uL *#Rnase Inhibitor 2uL *#M-MuLV reverse transcriptase 10 uL *Incubate the specimen with the reaction mixture for 15 min at 4Β°C, then transfer to 37Β°C for overnight.
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