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Does anyone have a workflow or knows of a kit that allows for whole transcript single cell RNA Seq with reverse transcriptase barcoding of each cDNA, and also UMI labeling? We are currently interested to perform this in our lab but most vendors are cutting down to 5' or 3' ends for scRNA-Seq analysis and we still want the whole transcript.
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NuGEN has a kit that has all the abovementioned features, the Ovation SoLo RNA-Seq Kit. -
I do not think this kit adds barcodes and UMIs to individual cDNA during RT as methods such as Drop-Seq does. Index (called barcode by NuGEN) and UMIs are added through adapter to amplified cDNA so it only identifies PCR duplicates generated during adapted library amplification. While Drop-Seq or 10X genomics single cell marks the 1st strand cDNA enabling identification of reads resulting from a single transcript.Originally posted by dwong View PostComment
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Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.07-01-2026, 11:43 AM
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I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.
Here are nine questions we think about, in roughly the order they matter, before...
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