Editing
Arichard:Notebook/2013/August
(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!
==== Oil and BSA incubation time test by qMDA ==== Motivation: Same as August 22. We need to find out why qMDA does not detect even the input amount, even after MDA, in microwells. Loss may be up to 90%. * Shearing due to glass pipets has been ruled out. * Shearing test was done on glass slides. We need to test PDMS arrays. * One possibility is that DNA is being removed with the oil. * Another is that DNA is binding to the well (in a time dependent manner), despite BSA coating. Objective: Test oil removal and DNA binding time as factors inhibiting detection by qMDA after extraction from PDMS microwell arrays. * 4 conditions: ** Load, extract immediately. ** Load, add 75 ul oil overlay, remove oil, extract immediately. ** Load, wait 1 hr, extract. ** Load, add 75 ul oil overlay, wait 1 hr, remove oil, extract. All conditions were loaded with 10 ul of 10 ng/ul genomic E. coli DNA. Conclusuion: 1 hr, no oil condition evaporated. Should have used coverslip. No oil, no wait condition had an excessive amount of DNA solution on top of the wells, which led to extraction volumes greater than 20 nanoliters (well volume) and lower CT values. The oil conditions did not show a difference between 1 hour and no wait. The DNA may be binding, but just very quickly. The wells still showed large loss of DNA. Note: 200 pg and 20 pg are on top of each other in the ladder. [[File: 2013_08_23 qmda oil and time test.xls| 2013_08_23 qmda oil and time test]]
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