Editing
Daniel:Notebook/HiResChrPaint/2013-7-12
(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!
==Research Notes: ULS== {| class="wikitable" |- align="center" | '''A''' || '''B''' |- | [[Image:ULS-LabelingRatio.png|400px]]|| [[Image:ULS-ReactionDuration.png|400px]] |- align="center" | '''C''' || '''D''' |- | [[Image:ULS-ReactionRatio.png|400px]]|| [[Image:ULS-ReactionTemp.png|400px]] |} Summary: '''A''': Optimal labeling ratio for ULS dyes is around 2%, or 1 in 50 bp. This experiment was done on Alexa 488 so it should be directly applicable to my research. '''B''': Increasing reaction time up to 1 hour should only improve labeling. Optimal time is suggested around 15 minutes but I think 20-30 wouldn't be too bad either. '''C''': Using more dye improves labeling efficiency, rounding off at around 1.4x dye:DNA. This is expected, but might have been an issue if the dye reacted with itself. '''D''': Optimal temperature for the reaction is between 70 and 80 C. Also expected, and not an issue in my protocols. Basically this tells me that as I expected, adding more dye and incubating longer should get me better yields. Also of note, reaction temperature is fine and I'm shooting for around 50 bp per dye molecule. The [[Daniel:Protocols/AlexaDyeCouple#ULYSISโ|ULYSIS Manual]] suggests a range of 43 to 67 bases per dye molecule, so this is about right. So my current thinking is that, as I suggested yesterday, I'm not getting all the dye that I want when I do my labeling reactions. Mabye the dye is sticking to the bottom of the tube or the sides, or in a giant clump. I'm not sure yet. But these results strengthen my suspicions from yesterday that I am not getting uniform dye concentration when I draw samples.
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