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hello, I would like to know if it is possible to check if in a rna-seq there are alternative splicing events, not comparing two condition but inside one condition only, that is if different transcript isoforms are produced in the same sample, thank you
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isoform detection
Check out Enzymatics AMP technology with individual cDNA molecule tagging. It will resolve your isoforms -
Cufflinks will compute the expression levels of individual isoforms.
Or, you can just examine the alignment files for individual genes in IGV (or the UCSC Genome Browser).
Obviously, it is not always possible to determine accurately to which transcript a read belongs, so the results are sometimes ambiguous.Comment
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cDNA tagging for isoform detection
Enzymatics AMP technology labels each cDNA with an adapter containing a unique random 8-mer sequence prior to amplification and NGS. This allows you to examine specific isoforms within the sequencing data. Check it out at www.enzymatics.comOriginally posted by blancha View PostComment
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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...06-18-2026, 07:11 AM -
Data variability is still an issue in sequencing technologies despite the advances in reproducibility and accuracy of these platforms. But the problem does not originate in the sequencing itself, but in the previous steps, before the sample reaches the sequencer.
The first step is collection, followed by preservation and sample preparation for analysis. Most scientists overlook those steps, but not being careful might just be skewing the experiment’s results.
...06-02-2026, 10:05 AM
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