I like to use the GETprime database, which gathers the sequences of optimized, isoform-specific qPCR primers. Check out the link.  

http://bbcftools.epfl.ch/getprime

Thanks Fabio. Appreciated. After a lot of effort with GAPDH i am thinking to change house keeping gene. I decided to go for B-actin. what you suggest? GAPDG is really a hell.

Hi Imran, 

Agreed! Have a look at this article regarding optimal housekeeping gene selection. The authors recommend beta-glucoronidase :

"In this study, the expression of a variety of housekeeping genes in a panel of 26 different human tumor and embryonal cell lines was assessed using real-time RT-PCR. For some control genes, the variability in expression was significant between different cell lines, despite the equalization of quantities of input RNA. The greatest variability was found for GAPDH. The lowest variability was found for β-glucuronidase (GUS) and 18S rRNA."

http://normalisation.gene-quantification.info/

Hello Fabio, Umair

Thanks for your suggestion. Actually i bought and tried both GAPDH and Actin as control. Luckily get very good results, Melting Curve Plot, Amplification cycle as well as Electrophoresis results all are fine now.

Really appreciate your concern.

Thanks