I am studying the expression pattern of few genes in tumor samples treated differently.

Following cDNA making and PCR i can clearly see difference in gene expression on gel. However even though i am using the same set of cDNA that i used for PCR i cannot detect significant fold change in expression of these genes in Real time quantification.

Everything is same in the conventional and real time PCR except the real time PCR has a different master mix containing the SYBR green and different polymerase.

Hence what can be the possible reasons for this? Any suggestions to troubleshoot this observation will be highly appreciated.

Thanks

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