Editing
Matt:LabNotes/2015-12-10
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!
=CircLigase II Reagent Testing= *Test new CircLigase II kit that arrived this month ==Reaction Mix== *5ul ssDNA control (10pmol total) *2ul 10X Reaction Buffer *1ul 50mM MnCl2 *4ul 5M Betaine *1ul CircLiase II Enzyme *7ul H2O *Total: 20ul *Sample1: New enzyme + kit reagents *Sample2: New enzyme + [[Hosuk:LabNotes/2014-3-24 | Hosuk's buffer 8-1-2014]] + Homemade MnCl2 & Betaine *Sample3: Old enzyme (red cap) + kit reagents *Sample4: No Enzyme ===Procedure=== #Incubated at 60C for 1hr #Incubated at 80C for 10min (inactivate) #Take 4ul of each and run TBU gel #ssDNA Zymo column purification #*Elute 10ul #Exonuclease I Digest half of sample #*5ul + 1ul 10X ExoIII Buffer + 4ul H2O #*5ul + 1ul 10X ExoIII Buffer + 1ul Exonuclease I + 3ul H2O #Incubate 37C for 45min #Incubate 80C for 15min (inactivate) #Run TBU gel [[File:2015-12-10_CircLigaseII_Test.jpg|650px]] ==Conclusion== *Both old and new enzyme work *Our homemade components work just as well *Circularized ssDNA actually runs slightly faster in our TBU gels
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