Editing
Sam:LabNotes/Micro-manipulation/2011-4-15
(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!
==Procedures== ===Test of pre-filtering the EtOH-fixed samples=== *Starting with EtOH fixed cell (stored in -80C) x2 This cell are about 100uL of OD600=1.0 E.coli, fixed in 500uL 100% EtOH for 3hr. #Pellet at 5min, 8000 rpm. Wash 1X in 1mL PBS(0.2um-filtered). ##One of the samples was filtered through 5um-filter, and labeled as "F". ##The other one was labeled as "R" following the regular procedure. #Resuspend in 0.5 mL PBS(0.2um-filtered). Transfer sample in a 1.7mL amber tube. #Prepare DAPI working solution (5mg/mL -> 1/5 dilution with H2O to 1mg/mL). #Add DAPI working solution in the E.coli ##Add 1uL dye for "R" tube. Add 4uL dye for the "F" tube. #Incubate in on the rocking plate/Eppendorf rocking incubator at RT (25C), 400rpm for 15min. ===Test of adding final washing with PBTw=== *Started from the DAPI-stained sample on 4-14-'11 #Separate the sample into two tubes(250uL/tube) #Pellet stained cells at 8000rpm, 5min, RT. #Prepare the PBTw (PBS containing 0.1% Tween20) by adding 40uL Tween20 into 40mL PBS(0.2um filtered). #Pick up one of the samples. Using 1mL PBTw to wash cells twice. Resuspend the cell pellet in 250uL PBTw. Label the tube as "washed". #Label the other tube as "non-washed".
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