I would like to know the cDNA quality before proceeding to PCR. I learnt the use of Nano drop may not be helpful as the cDNA synthesis makes use of RNA and mixture of dNTPs, random hexamers and/or oligo-dT which will influence the A260/A280 ratio.
Thanks!
Hi Godson Aryee,
The best way to find out is to do a PCR:
Nice specific bands for conventional PCR or early CTs for real-time PCR.
Hi Godson Aryee
You can also do PCR by exon-intron spanning primer to verify the gDNA contamination on your cDNA product.
Thank you
Hi Godson Aryee,
the most convenient method to check the quality would be an exon-spanning PCR for a housekeeping gene (HKG) as mentioned. If the quality is ok, than you already have the HKG data which you always need for PCR analyses. For endpoint PCR I would prefer a larger PCR product (200-500bp), for qPCR a smaller (
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