3 replies to this topic

[connicelee]

hi all,

I will start to run real time PCR using SYBR green to quantify bacteria. My sample will be genomic DNA. I am not sure whether to use genomic DNA or plasmid DNA (which i had clone the target gene in)as my standard curve. However if genomic DNA is used, i don't have the information of the genome size. How can i calculate the copy number? Or i have to use kit to quantify it?

TQ.


Regards,
Connice

Edited by connicelee, 12 December 2004 - 09:34 PM.

[nabla]

Plasmid sound good. Better would be DNA because it is your template for this PCR (kinetic). Estimating the DNA-size would probably be too unexact?!

[pcthanh82]

Follow me. You first design a primer then run PCR.

[connicelee]

Thanks. ..... the primers was designed using primer express 2 and now synthesizing..........because of the hilodays........i have to wait untill next year.

If i use the genomic dna as my standard curve,i have to know the genome size to calculate the copies number. My problem is i do not have the information about the genome size. Should i use picco green double stranded dna quantification kit to quantify the copy number of the genomic dna?


TQ......

Regards,
Connice