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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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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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