Editing
Hosuk:LabNotes/2014-5-6
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]] ===Test : +/- a-dUTP, and long RT incubation at 4C=== ====1st Try==== *8 MatTek dishes - PGP1F (05/05) **10% Formalin for 15min at RT(25C), **0.25% Triton X-100 for 10min at RT **0.1N HCL incubation at RT *BUT! Samples were gone, destroyed! **upto 0.1N HCL process seemed too harsh **Letβs just do Triton X-100 and run reverse transcript! ====2nd Try==== *With only 4 sample (05/06) **Sample 1 : +a-dUTP, Random hexamer, 15min incubation at 4C **Sample 2 : -a-dUTP, Random hexamer, 15min incubation at 4C **Sample 3 : +a-dUTP, ACTB, RAB7A primer, 1hr. incubation at 4C **Sample 4 : -a-dUTP, ACTB, RAB7A primer, 1hr. incubation at 4C ====Procedure==== *Follow the latest protocol (Nature protocol manuscript) *#10% Formalin for 15min at RT(25C) --> done at CERC *#Wash with PBS 3 times --> done at CERC *#Move samples to the lab at PFBH *#Add 0.25% Triton X-100 in DEPC-PBS and incubate for 10min at room temp. *#mean while prepare RT mix and put the mix on an ice *#Aspirate 0.25% Triton X-100 in DEPC-PBS and wash with PBS 3 times *#Add RT mix and put in 4C, incubate for 15min. (1hr. for other 2 sample of target specific primers) *#Move samples to 37C and incubate for 12hr.
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