Can i use genomic dna for the preparation of standard curve for real time pcr?

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Erika Feltrin

Hi Jeny,

it depends what kind of qPCR you are doing? Sybr or TaqMan? Which primers are you using?

Erika

Govinda Sharma

How much genomic DNA are you inputting per reaction? Amplification becomes inhibited dramatically the more gDNA you add. I try to avoid going above 10ng in a 10uL reaction volume.

Chongtham Rajiv

Yes you can use.

First amplify the 10ng-60ng genomic DNA with the primer to be use for quantification

Check for amplification in agarose gel, if sharp band is observed proceed for further step.

Clean up the PCR product using PCR cleanup kit.

Quantified using spectrophotometer method like Nano-drop.

Create a serial dilution of desire range for standard and use for quantification.

I have done same and got the efficiency around 100%.