Dilute DNA concentration - (May/19/2007 )

Dear All, I need to test the sensitivity of my PCR.
Have anybody dilute the extract DNA in varie concentrations ? Is it necessary to incubate in 50-60 degree celsius before dilute the DNA (someone tell me like that)?
Alternatively, Could I dilute my bacterial suspension in varie concentration before extraction the DNA ?
Help me, please.

-krapom-

I'm guessing you want to try different concentrations of plasmid DNA as a template for PCR?

Just dilute it in TE (or water). No need to incubate or anything.

QUOTE

Alternatively, Could I dilute my bacterial suspension in varie concentration before extraction the DNA ?

I wouldn't do that, you'd never be able to predict the concentration, and it's WAY more work than just diluting the DNA!

-Zouden-

QUOTE (Zouden @ May 19 2007, 05:33 PM)

I'm guessing you want to try different concentrations of plasmid DNA as a template for PCR?

Just dilute it in TE (or water). No need to incubate or anything.

QUOTE

Alternatively, Could I dilute my bacterial suspension in varie concentration before extraction the DNA ?

I wouldn't do that, you'd never be able to predict the concentration, and it's WAY more work than just diluting the DNA!

Thank you for your kindness. I need to set my PCR testing. I try to diltue my genomic DNA in varie concentration and then test my PCR. A doctor told me to incubate the DNA in every steps of dilution. I think it is nonsense and spend much time to do

I will follow with your sugeestion.

-krapom-