Editing
Chromosome Separation Attempt 2 (2/25/10)
(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!
==Preparation of Isolation Buffer== Solutions of 100mM MgSO4, 55mM KCl, 5.5mM HEPES, and 120mM DTT were needed. For each (minus the DTT), the number of moles per gram of solid was calculated and the resultant molarity from adding 1mL water was then found. Using this as a starting point, the actual amount of water needed to create solutions of the correct molarity was calculated. For MgSO4: **Insert calculation** Similarly, for KCl: Add 0.4g KCL (solid) to 48.764mL water -> 110mM KCl solution For HEPES: Add 0.4g HEPES (solid) to 152.58mL water -> 11mM HEPES solution Because the stock solution of DTT was far more concentrated (1 M) than the needed concentration, a smaller total volume of DTT was added to the isolation buffer, rather than adding the full 0.25mL of diluted DTT. The combined solution of 55mM KCl and 5.5mM HEPES was created by adding 4.5mL 110mM KCl and 4.5mL 11mM HEPES to a separate container, thereby reducing the concentrations in half, but allowing for a total volume of 9mL In the end, the buffer was made as follows: 1mL 100mM MgSO4 9mL 55mM KCl + 5.5mM HEPES 0.03mL 1M DTT 0.7mL 10000U/mL RNase The solution was then filtered via a 10mL syringe with a 22um filter attached. The filtration rate was slow - drop by drop - as to be expected from a filter.
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