Editing
Sam:LabNotes/Human/2009-3-24 exp2
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!
='''High through-put single gene PCR validation with template dilution on 1/25, 1/50, 1/100'''= ==Procedure== *Primer dilution x 8 (primer 1-1~1-8 from set-1) 1 rxn 20 rxn Primer (f+r) 0.5 10 uL H2O 3.5 70 uL ------------------------------------ 4.0 80 uL *Aliquots Taq2X enzymes into PCR tubes (100 uL) x 8 PCR tubes *Exp design Row A: B7 - 1/25(from exp of test2 2-25-2009), amplicon from whole single cell Row B: B7 - 1/50 Row C: B7 - 1/100 Row D: D5 - 1/25(from exp of test 2-27-2009), amplicon from 20 cells. Row E: D5 - 1/50 Row F: D5 - 1/100 Each template was tested on eight primers (1-1 ~ 8-1) *Amplication: Program: T52C30 (Tm=52C, for 30 cycles) *Gel electrophoresis **2% agarose gel, midium tray with 26-well comb **Sample (9uL) + Loading buffer (2 uL) **Run at 135V for 20 min ==Results== [[Image:ZhangLab_2 2009-03-24 18hr 58min-annotate.jpg|700px]] [[Image:ZhangLab_2 2009-03-25 10hr 51min-annotate.jpg|700px]] Primer(amplicon size) 1-1(270), 2-1(540), 3-1(235), 4-1(383), 5-1(413), 6-1(299), 7-1(242), 8-1(230) ==Discussion== *The dilution of template (from original MDA amplicon ~2000 ng/uL) into 1/25, 1/50 or 1/100 didn't affact the appearance of bands in the current experiment. *Amplication of 30 cycles improved the band intensity obviously. *Gel electrophoresis ran at 20 min is optimal for this screening.
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