Editing
Hosuk:LabNotes/2013-11-18
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]] ===Experimental Plan=== ====Rolony in MatTek dish, 96 well plate==== *MatTek dish **Fix : **Aspirate media, Add 10% Formalin, Incubate 37C for 15min. **Wash with PBS 3 time, Add 2mL Ethanol *96 well plate **In T25 flask, Add 1mL Trypsin, Incubate 37C for 5min. **Add 4mL media (Total 5mL), Take 50uL, add in each well **Fix by aspirating media, adding 10% Formalin *Check cells after Fixation ====Dye labeled Probe Annealing Test==== *Check Non-specific binding, and Upper limit **Use dcProbe#-Cy3 --> check non-specific binding **Use Cy5-1st Rolony --> Upper limit *Dye-Probe mixed with different concentration of Formamide **10%, 30%, 50%, 70% in 2x SSC *Incubating temperature **?? *Check the number of Rolonies ====Padlock Probe Concentration Test==== *Lower limit, Upper limit of the concentration *Normally 0.2uM for each Padlock Probe after PCR *Use One MatTek Dish *Use ppMALAT1 **0.1uM, 5uM, and check the signal, then test middle concentration *Preheat ppMALAT1 at 80C *Incubate at 50C for 15min ====Ampligase test==== *Use one MatTek dish *Add Padlock probes, Ligase, 2nd RCA and check how many 2nd Rolonies *Ampligase reaction condition **Add 3 Padlock probes β ppACTB, ppRAB7A, ppMALAT1 **Padlock probe concentration : 1uM each (? could be changed) **Padlock probe incubation time : 30min **Ampligase concentration : 5U **Temperature : 60C **Incubation time : 4hr. [[File:Plan_2013-11-18.png|400px]]
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