Editing
Andy Chen:Micromanipulation
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!
=Micro-manipulation Technique Notes= ==DNA Extraction from Microwells After MDA== ===General Protocol=== *Prepare pipette tips, tubes for collected samples, and a tube of medium (PBS, H2O, etc). UV everything 10-15 mins. *Transfer desired amount of medium to each sample tube (for example, fill tube with 1 μL if doing a subsequent 20 μL MDA amplification) *Load a tip into the pipetter and pre-load the tip with medium. To do this, move the pressure to slightly positive (+10 to +20), immerse the tip in the tube of medium, move the pressure to slightly negative (-5 to -10), wait for the pressure to equalize (it will stop clicking), then move the pressure back to zero. *Move the tip to the desired well. Take a picture of the fluorescence for reference. *Apply a positive pressure to eject some of the fluid into the well (enough to cover the bottom of the well). Move the tip around to help loosen the DNA-bead complex. Apply a negative pressure to extract the DNA. Check fluorescence to see if DNA was extracted adequately. If not, repeat ejection-extraction a few more times to collect as much DNA as possible. *Move tip to home and eject the collected DNA into sample tube by first applying a large positive pressure (+50, for example) and then hold the blue [INJECT] button for a few seconds to insure all the DNA was ejected. ===Tips=== *Tips around 20-50 μm in diameter are ideal. *When filling a dry microwell with medium, there seems to be a threshold to getting the medium out of the tip. Thus positive pressure needs to be applied carefully to prevent flooding and possible contamination. Pressure should be increased slowly. If there appears to be a bulge of medium on the tip, try to move the tip around on the bottom of the well to get some liquid into the well, making it easier to add medium. *I found it easier to move to the general location of the well using the [0] setting, then changing it to [1] for finer movements. [0] seems to be affected by acceleration, so if you aren't careful, it is easy to over move it and smash the tip into the well. ==Single-cell Extraction== ===General Protocol=== {|align=right |[[File:slide.gif|align=right]] |} *Prepare tips, sample tubes, medium tubes, tubes for diluting cells (if necessary), slide and coverslip. UV everything 10-15 minutes. *
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