Hello,

I am using Superscript IV first strand synthesis kit for making cDNA from RNA samples. The 260/280 and 260/230 values of My RNA samples are above 2. However after reverse transcription with superscript IV kit (invitrogen) I got really poor quality in terms of concentration and 260/280 values. For cDNA, the 260/280 values should be 1.80 or more but I am getting lesser than that. The 260/230 values are < 1.4. Previously, I used to use the High capacity Reverse transcription kit (Applied biosystem) and never had this issue.

I am making cDNA from 1 ug and 3 ug RNA as the superscript IV claims to make cDNA from 20 pg to 5 ug of RNA. But with 3 ug of RNA. I am not getting higher concentration of cDNA. Also, the quality is not good for both 1 ug and 3 ug. When I tested the same RNA samples with High capacity cDNA kit from Applied biosystem, I am getting good result. The problem is I have to use the superscript IV kit as I am planning to make more concentrated cDNA. Is there anyone who work with superscript IV kit? I wanted to know their experience with it. It's an urgent issue.

Thanks.