You can scale up reverse transcription reactions with no issue; using 3 µg of input RNA should be fine as long as you increase the other components as well.

thanks for your answer.

Is it possible in your opinion to use 3 ug of RNA without increasing the amount of the other cimponents? I'm trying to incrase the concentration, so this solution is not optimal.

thanks,

Sapir

You can probably increase the amount RNA input for your reverse transcriptions above the recommended amount, but this may saturate the reverse transcriptases leading to increased amounts of unconverted RNA.

If you need to increase your cDNA concentration to such high levels in order to detect your target transcript, it might be advisable to use gene/transcript-specific primers for the reverse transcription rather than using oligo-d(t) or random hexamers.

I always put more RNA for RT. should be OK.

Just a suggestion, probably you may scale up all reagents 3-5 times including your RNA and then just go through ethanol precipitation after RT, and elute your sample in smaller volume of appropriate buffer. That would give you higher concentration and purer cDNA samples.

thank you for your answers.

Sapir