Editing
Daniel:Notebook/ComboLock/2017-2-3
(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!
==Protocol-Part 4-RCA== <ol start="8"> <li>Exonuclease Digestion</li> <ol type="A"> <li>Mix 15 uL Exo I (20U/uL) and 3 uL ExoIII (100U/uL)</li> <li>Add 2 uL to each sample, mix by swirling pipette tip</li> <li>Incubate for 1 hour at 37C</li> <li>Heat kill by incubating for 5 min at 95C</li> </ol> <li>Rolling Circle Amplification</li> <ol type="A"> <li>Prepare 8.2X Master mix according to table below; '''Give A samples LLRC and B Samples LLRC*'''</li> {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |- style="background-color:#92CDDC;font-size:12pt;font-weight:bold" align="center" | width="155" height="30" | Reagent | width="65" | uL Added | width="105" | Master Mix (8.2X) | width="105" | Master Mix (4.2X) |- style="font-size:12pt" | height="15" valign="bottom" | Template | align="center" align="center" valign="bottom" | 5 | align="center" valign="bottom" | NA | align="center" valign="bottom" | NA |- style="background-color:#BFBFBF;font-size:12pt" | height="15" valign="bottom" | RCA Primer (10 uM) | align="center" align="center" valign="bottom" | 2.5 | align="center" align="center" valign="bottom" | 0 | align="center" align="center" valign="bottom" | 0 |- style="font-size:12pt" | height="15" valign="bottom" | dNTP (1 mM) | align="center" align="center" valign="bottom" | 5 | align="center" align="center" valign="bottom" | 41 | align="center" align="center" valign="bottom" | 21 |- style="background-color:#BFBFBF;font-size:12pt" | height="15" valign="bottom" | 10X Buffer | align="center" align="center" valign="bottom" | 2 | align="center" align="center" valign="bottom" | 16.4 | align="center" align="center" valign="bottom" | 8.4 |- style="font-size:12pt" | height="15" valign="bottom" | Phi29 | align="center" align="center" valign="bottom" | 1 | align="center" align="center" valign="bottom" | 0 | align="center" align="center" valign="bottom" | 0 |- style="background-color:#BFBFBF;font-size:12pt" | height="15" valign="bottom" | BSA (10 mg/mL) | align="center" align="center" valign="bottom" | 0.4 | align="center" align="center" valign="bottom" | 3.28 | align="center" align="center" valign="bottom" | 0 |- style="font-size:12pt" | height="15" valign="bottom" | nfH2O | align="center" align="center" valign="bottom" | 4.1 | align="center" align="center" valign="bottom" | 33.62 | align="center" align="center" valign="bottom" | 18.9 |- style="background-color:#BFBFBF;font-size:12pt" | height="15" valign="bottom" | Total |style="font-weight:bold" align="center" align="center" valign="bottom" | 20 |style="font-weight:bold" align="center" align="center" valign="bottom" | 114.8 |style="font-weight:bold" align="center" align="center" valign="bottom" | 58.8 |} <li>Add 11.5 uL master mix to each tube</li> <li>Add sample and 1 uL appropriate enzyme to new reaction tubes</li> {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |- style="background-color:#B1A0C7;font-size:12pt;font-weight:bold" align="center" | width="100" height="30" | Sample | width="65" | 1X (AB) | width="65" | 2X (AB) | width="65" | 3X (AB) | width="65" | 4X (AB) | width="65" | 1Y (AB) | width="65" | 4Y (AB) |- style="font-size:12pt" | height="15" valign="bottom" | Phi29 Source | align="center" valign="bottom" | NEB | align="center" valign="bottom" | NEB | align="center" valign="bottom" | NEB | align="center" valign="bottom" | NEB |style="font-weight:bold" align="center" valign="bottom" | Epicentre |style="font-weight:bold" align="center" valign="bottom" | Epicentre |} <li>Incubate at 37C for 3 hours</li> <li>Incubate at 65C for 10 minutes;Hold at 10C until next step</li></ol></ol>
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