Hi everyone,

First of all, thank you for attention and answers. I did reverse transcriptase PCR for obtain cDNA from RNA. I used 3 different cell line RNA and my results looks fine. Approximately, all concentrations are 1900-2000 ng/uL and A260/280 value is 1,80-1,83.

My problem is that my controls. I used Revert aid first strand cDNA synthesis kit. And according to the kit protocol, I have 2 negative control which are reverse transcriptase neg. and GAPDH RNA template neg., respectively. Also, I measured them on the nanodrop ssDNA and dsDNA. And the results are in the below:

RT neg ssDNA measurement - 1903,6 ng/uL, 1,84 A260/280, 1,98 A260/230.

RT neg dsDNA measurement - 2884,0 ng/uL, 1,85 A260/280, 1,98 A260/230.

RNA template neg ssDNA measurement - 1903,1 ng/uL, 1,83 A260/280, 1,95 A260/230.

RNA template neg dsDNA measurement - 2889,7 ng/ul, 1,83 A260/280, 1,94 A260/230.

Due to they are negative control, their measurement should be close to 0.00? I will do PCR later with primers for checking but these results have me concerned.