Editing
Hosuk:LabNotes/2014-8-7
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]] ===Cancer Mutation detection=== ====Sample==== *Sample **NCIH1975_S1_2014-08-05 : -dUTP, rHex ο 45C **NCIH1975_S2_2014-08-03 : +dUTP, rHex ο 60C --> will use this later! ====Run Ampligase + Klentaq and 2nd RCA==== *Reaction Mix [[File:Ampligase_Reagent_2014-08-07.jpg|600px]] *Start Ampligase+Klentaq : 08/05, 3:30pm (Just use NCIH1975_S1_2014-08-05), ~21.5hr at 45C *Start RCA : 08/06, 2:40pm **1uM ppRCA in 200uL 2x SSC, add and incubate at 42C for 36min, **Wash with 2x SSC twice, 1x SSC twice, 1x PBS twice, then add Phi29 mix *Finish : 08/07 (RCA for ~18.5hr) ====Decoding==== *Decoding β 08/07 *Sample : NCIH1975_S1_2014-08-05 : -dUTP, rHex *1st Rolony β Cy3 : 1uM *dcProbe0~4 : 488, Cy3 : 1uM **dcProbe[#]-488 and dcProbe[#]-Cy3 are mixed in a sampe 100uL tube *Step *#Add dcProbe[#], incubate for 10min, image *#Strip with 80% formamide(pre-heated at 70C) for 15min, wash with PBS twice *#Repeat applying dcProbe and imaging *Result : under anaylzinging
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