Editing
Alice:LabNotes/2010-4-2
(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!
===Step 5. Wash the Streptavidin Dynabeads Plus Bound DNA=== #After the 45-minute incubation, transfer the entire content of each 0.2ml tube to a 1.5ml tube. #Use the magnet from the DynaMag-2 device to bind the beads. Remove and discard the liquid once clear. #Add 100μl of SC Wash Buffer I heated to 47°C. Mix by vortexing for 10 seconds. #Place the tubes in the DynaMag-2 device to bind the beads. Remove and discard the liquid once clear. #Remove the tubes from the DynaMag-2 device and add 200μl of Stringent Wash Buffer heated to 47°C. Pipette up and down 10 times to mix. Work quickly so that the temperature does not drop much below 47°C. #Incubate at 47°C for 5 minutes. #Repeat Steps 5.5 - 5.7 for a total of 2 washes with Stringent Wash Buffer heated to 47°C. #Place the tubes in the DynaMag-2 device to bind the beads. Remove and discard the liquid once clear. #Add 200μl of room temperature SC Wash Buffer I and mix by vortexing for 2 minutes. If liquid has collected in the tube’s cap, tap the tube gently to collect the liquid into the tube’s bottom before continuing to the next step. #Place the tubes in the DynaMag-2 device to bind the beads. Remove and discard the liquid once clear. #Add 200μl of room temperature SC Wash Buffer II and mix by vortexing for 1 minute. #Place the tubes in the DynaMag-2 device to bind the beads. Remove and discard the liquid once clear. #Add 200μl of room temperature SC Wash Buffer III and mix by vortexing for 30 seconds. #Place the tubes in the DynaMag-2 device to bind the beads. Remove and discard the liquid once clear. #Remove the tubes from the DynaMag-2 device and add 50μl PCR grade water to each tube of bead-bound captured sample. #Store the beads plus captured samples at -15°C to -25°C or proceed to Chapter 7, Captured DNA Amplification Using LM-PCR. *Note: There is no need to elute DNA off the beads. The beads plus captured DNA will be used as template in the LM-PCR.
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