RNA was extracted from micro-dissected (under microscope) epithelial layer of the tissue as described earlier. mRNAs were isolated from the total RNA with a commercially available kit (Dynabeads® mRNA DIRECTTM Micro Purification Kit, Life Technologies). This was followed by cDNA Library construction using Ion Torrent RNA-Seq Kit (Life Technologies) for whole transcriptome libraries. For individual sample Barcodes 1 through 8 were attached using Ion Xpress 1–16 barcoding kits. Quantitation of cDNA libraries was performed using the Ion Library Quantitation Kit (Life Technologies) to evaluate appropriate template dilution factor for subsequent emulsion PCR and sequencing. This was followed by next generation sequencing using the Ion Torrent platform according to manufacturer’s protocols ( Life Technologies, Carlsbad, CA)
Dynabeads mrna directtm micro purification kit
Manufactured by Thermo Fisher Scientific
The Dynabeads® mRNA DIRECTTM Micro Purification Kit is a product designed for the isolation and purification of mRNA from small samples. It utilizes magnetic beads coated with oligo(dT) to capture and extract mRNA directly from cell lysates or tissue samples.
Automatically generated - may contain errors
2 protocols using dynabeads mrna directtm micro purification kit
1
Transcriptome Analysis from Micro-dissected Tissue
2
Transcriptome Analysis of Micro-Dissected Tissue
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RNA was extracted from micro‐dissected (under microscope) epithelial layer of the tissue as described earlier. mRNAs were isolated from the total RNA with a commercially available kit (Dynabeads® mRNA DIRECTTM Micro Purification Kit, Life Technologies). This was followed by cDNA library construction using Ion Torrent RNA‐Seq Kit (Life Technologies) for whole transcriptome libraries. For individual sample, barcodes 1‐8 were attached using Ion Xpress 1‐16 barcoding kits. Quantitation of cDNA libraries was performed using the Ion Library Quantitation Kit (Life Technologies) to evaluate appropriate template dilution factor for subsequent emulsion PCR and sequencing. This was followed by next‐generation sequencing using the Ion Torrent platform according to manufacturer's protocols ( Life Technologies, Carlsbad, CA).
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