Editing
Daniel:Protocols/FISH
(section)
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!
===Hybridization=== ''Original Protocol from Beliveau et al'': [[Media:Interphase_FISH_protocol.pdf|FISH Protocol (Beliveau)]] ''Day 1'' #Allow stored slides to warm to room temperature #Incubate slides in 2X SSCT +50% (v/v) formamide for 2.5 minutes at 92 C in a pre-warmed coplin jar #Incubate in coplin jar containing 2X SSCT + 50% formamide at 60C for 20 minutes #Remove slides and allow to cool to room temperature #Add 25 uL hybridization cocktail to a 22x22 #1.5 coverslip #Invert slides on to cocktail-covered coverslips; seal with rubber cement #Allow the rubber cement to air-dry for 5 minutes at room temperature #Denature for 2.5 minutes at 92C; keep humid #Transfer slides to a humidified chamber and hybridize overnight at 37C or 42C ''Day 2'' #The next day, remove coverslip carefully and wash slides in pre-warmed jar containing 2X SSCT at 60C for 15 minutes #Incubate slides in 2X SSCT at room temperature for 10 minutes #Incubate slides in 0.2X SSC at room temperature for 10 minutes #Tap slides dry on papertowels; do not let the surface containing the cells touch the paper towels #Add 15 uL mounting media such as SlowFadeGold with DAPI to a 22x30 #1.5 coverslip #Invert slides onto coverslips with mounting media #Seal each slide using nail polish #Allow at least 30 minutes for nail polish to dry #Slides are quite stable for several months at 4C
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