Editing
AlanFung:Protocol/Construction of Solexa sequencing Library
(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!
===PCR of sequencing library=== Solexa_PCR_up: AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT AmpR6.3Sol: CAAGCAGAAGACGGCATACGAGCTCTTCGGAACGATGAGCCTCCAAC AmpF6.3rSol: CAAGCAGAAGACGGCATACGAGCTCTTCCAGATGTTATCGAGGTCCGA {| {{table}} | Ligation products||10||10 |- | 10uM solexa PCR up||2||2 |- | 10uM AmpR6.3Sol||2||- |- | 10uM AmpF6.3Sol||-||2 |- | 2X iProof master mix (Bio-Rad)||50||50 |- | 50X SYBR Green I||0.4||0.4 |- | H2O||36||36 |- | |} *Setup 2 master mix {| {{table}} | ||F||R |- | Ligation products|||| |- | 10uM solexa PCR up||13.2||13.2 |- | 10uM AmpR6.3Sol||13.2||- |- | 10uM AmpF6.3Sol||-||13.2 |- | 2X iProof master mix (Bio-Rad)||330||330 |- | 50X SYBR Green I||2.64||2.64 |- | H2O||237.6||237.6 |- | |} PCR program: 98 °C 30sec -> 8 cycles of (98°C 10sec -> 60°C 20 sec -> 72°C 15sec) -> 72°C 3min ->15°C hold. [[Image:20091112_125359.jpg]] * TBE Gel verification (10well) * 15ul h2o+4.5ul 6x loading dye+0.5ul 25bp ladder * 15ul h2o+3ul 6X loading dye+2ul sample [[Image:ZhangLab_2 2009-11-12 14hr 46min.jpg]] [[Image:ZhangLab_2 2009-11-12 14hr 48min.jpg]] *Mix the amplicons with two sets of primers, purified with Qiaquick columns with 32ul EB buffer
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