Editing
Kun:LabNotes/ASE/2007-11-26
(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!
==Exp.2 Test the protocol of quantifying DNA with PicoGreen on Chromo4== #Make 500ul of 1:200 diluted PicoGreen: 2.5ul PicoGreen + 497.5ul 1X TE. #Make 100ul 2ng/ul lambda DNA solution: 2ul 100ng/ul stock DNA + 98ul 1X TE. #Make serial dilution of DNA standard: 1X TE 2ng/ul DNA Diluted PicoGreen Total volume 0ul 25ul 25ul 50ul 20ul 5ul 25ul 50ul 22.5ul 2.5ul 25ul 50ul 24ul 1ul 25ul 50ul Get fluorescent reads on Chromo4 on row 1-4, two times per position. {| {{table}} | align="center" style="background:#f0f0f0;"|''' ''' | align="center" style="background:#f0f0f0;"|'''Conc. (ng/ul)''' | align="center" style="background:#f0f0f0;"|'''1.1''' | align="center" style="background:#f0f0f0;"|'''1.2''' | align="center" style="background:#f0f0f0;"|'''2.1''' | align="center" style="background:#f0f0f0;"|'''2.2''' | align="center" style="background:#f0f0f0;"|'''3.1''' | align="center" style="background:#f0f0f0;"|'''3.2''' | align="center" style="background:#f0f0f0;"|'''4.2''' | align="center" style="background:#f0f0f0;"|'''4.1''' |- | A||1||3.43||3.45||3.62||3.63||3.72||3.69||3.37||3.41 |- | B||0.2||0.52||0.52||0.51||0.51||0.53||0.53||0.51||0.52 |- | C||0.1||0.34||0.33||0.32||0.32||0.33||0.32||0.32||0.33 |- | D||0.04||0.2||0.2||0.19||0.19||0.2||0.2||0.22||0.22 |- | E||1||2.05||2.06||2.06||2.06||2.17||2.16||2.14||2.14 |- | F||0.2||0.51||0.51||0.52||0.52||0.52||0.52||0.56||0.53 |- | G||0.1||0.33||0.33||0.32||0.32||0.34||0.33||0.33||0.33 |- | H||0.04||0.2||0.2||0.2||0.2||0.19||0.2||0.2||0.2 |- | |} Scatter plot: X: concentration (ng/ul); Y: fluorescent intensity [[Image:2007-11-26-picotest-xyplot.png|400px]] <b>Conclusion:</b> I think Chromo4 did get linear read-outs on PicoGreen stained DNA. This could be used to measure DNA concentration. However, a ssDNA standard should probably be used for ssDNA samples. I measured the concentration of the eight Solexa libraries using this method. However the concentration seems to be lower than what I expected. Perhaps it is due to the fact that I exposed the picogreen to light (and probably UV) for a while. {| {{table}} | align="center" style="background:#f0f0f0;"|'''Library''' | align="center" style="background:#f0f0f0;"|'''ng/ul''' | align="center" style="background:#f0f0f0;"|'''corrected,ng/ul''' |- | Hues6 gDNA||0.53||1.07 |- | Hues6 cDNA||1.34||2.69 |- | H1 gDNA||1.59||3.17 |- | H1 cDNA||1.97||3.94 |- | H2 gDNA||1.66||3.32 |- | H2 cDNA||0.98||1.96 |- | BJ MDA DNA||0.18||0.36 |- | BJ cDNA||0.89||1.78 |- | |} <b>Note: I just realized that the concentrations should be multiplied by two, and then they are fairly close to what I got from the PAGE quantification.</b>
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