I am trying to do dscDNA from Immortalized Fibroblasts using Illumina kit. RNA is extract with trizol, purified with QIAgen column, and treated with DNase I. We found no degradation and no contamination (Nanodrop and Agilent) of the RNA. Then we try to do fs and dscDNA and check new synthetized DNA with nanodrop. Profiles are awfull . We are using the same protocol for Immortalized Lymphoblasts and it's working very well.
We tried differents concentration of RNA, but it didn't worked.
SO if anybody has an idea of how I could do it will be very helpfull !!
THanks a lot in advance !
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ds cDNA synthesis problem
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