Rt pcr - (Jul/09/2015 )

I have isolated RNA from HEK cell lines.While doing one step RT PCR using kit, and then running the gels, the negative control also shows a band at the same size as of desired for the sample.Why am I getting a band in negative controls?

-shubhitamathur-

Most likely due to contamination in one of your reaction component.

What about your positive controls?

-Mad Researcher-

Positive control also shows the respective band.The main problem is that why the band is observed in negative controls.I took all the precautions to set the reaction.There is a separate RNA working table.Still,there is a band in negative controls??

-shubhitamathur-

What is your negative control? Could it be showing DNA contamination in your prep?

-almost a doctor-

Negative control is cells only.....i.e cells in which no transfection took place.When there is no transcription,how can the desired band of 300 bp shows up there?

-shubhitamathur-