Using cDNA as input for TruSeq: Anyone tried this?

[sweetph3]

We have a cDNA sample that we would like to use to make a TruSeq library. We tried Nextera already, but Nextera due to how it tagments doesn't give us all of the 5' sequence. The cDNA is already ~400bp-2kb so we don't want to fragment it. We tried using ~2ng as input into the TruSeq prep starting at the "Perform End Repair" step. The library prep failed. Theoretically, this should work, correct? Has anyone tried doing this? If so, how much cDNA input (or any other dsDNA, like PCR product) did you use?

Thanks for any suggestions!

ETA: We normally do TruSeq RNA kits, so I apologize for my ignorance on this topic. I see that the TruSeq DNA kits use more input than I did (100-200 ng for TruSeq DNA Nano; 1-2 ug for TruSeq PCR-free). I'll try again using more input, but would still appreciate anyone's suggestions!

[kwaraska]

TruSeq works on cDNA. Just treat it as DNA.

[frozenlyse]

Truseq works on double stranded DNA - if your cDNA is single stranded, you will have to synthesise a second strand before doing the 'end repair' ie end blunting then a tailing.