Editing
Sam:LabNotes/Microbiome-new/2009-2-20
(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== ==='''E. coli enrichment and dilution'''=== *Warm up the LB broth to RT (sterile, storedin 4 degree fridge). Prepare two 250 mL flask and filled with 50 mL LB broth. Inoculate 50 uL of E. coli in 50 mL LB both (1:1000 ratio). *Mix the broth and cap the flask-top with foil. Incube the flask in 37-degree incubator overnight. *The next day. Harvest the enriched E. coli by transferring 2 mL culture into 2-mL microcentrifuge tubes. Prepare 8 tubes and centrifuge at RT, 4000 rpm for 5 min to pellet the bacteria. *Remove the supernant. Perform PBS washing (using 1 mL PBS) and concentrating the pellet during each PBS washing *Measure the optical density using Nanodrop (chose 'cell-cultur' function, measure 1.5 uL E. coli suspension using OD600 (default) and OD590 (custom) nM wavelength. *Make sure to blank Nanodrop using PBS (the media used in BAC dilution). Try to adjust the O.D600 = 1.0, which should give a bacteria number around 10^9 CFU/mL = 10^6 CFU/uL *Dilute the BAC in series dilution as following: Dilution Factor Start 1/100 1/1000 1/10000 1/100000 -------------------------------------------------------------------- BAC mixture 10 uL 10 uL 10 uL 10 uL 10 uL PBS 990 uL 90 uL 90 uL 90 uL -------------------------------------------------------------------- Concentration 10^6/uL 10^4/uL 10^3/uL 10^2/uL 10/uL ** Mix by repeat pipetting using p200 pipettor in each step ==='''Bacteria MAD reaction'''===
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