DNA-Standards for quantitative PCR

tim tim at besancon.net
Wed Mar 24 08:36:26 EST 1999


Using native or linearised Plasmid for standards ?
No difference : we have tested the efficiency of linearized standard
versus native.

How do you produce your linearised Standard ?
Digestion
(We tried gel elution using QuiaExII, which seem'd to work)
Do you recommend Carrier DNA and which type ? (We used Salmon Sperm DNA
from Stratagene which had a profound negative effect on PCR-reaction)
Which method of DNA-Quantification do you prefer ? (What about your
experience with OD at 260 nm compared to other, more sophisticated
methods?)
How about long-term storage of your standards ? Does this work, or do
you
prefere to produce a new set of standards after a period of time ?
Thank you very much.






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