We are pleased to report the first customer poster featuring the newest addition to our product lineup, the SageELF whole-sample fractionation tool. “Single Molecule, Real-Time Sequencing of Full-length cDNA Transcripts Uncovers Novel Alternatively Spliced Isoforms” comes from scientists at Pacific Biosciences and the University of Washington.
The poster describes a human gene expression study in which the scientists were able to generate full-length isoform sequences, with some transcripts longer than 10 Kb. Isoform sequencing was conducted using the PacBio® RS II DNA Sequencing System. The cDNA sequences came from the MCF-7 human breast cancer cell line as well as from brain, heart, and liver cells.
The authors note: “Even in extensively profiled sample types, the method has been able to uncover large numbers of novel alternatively spliced isoforms and previously unannotated genes.” This ability to produce full-length transcript sequences offers a unique way to examine alternative splicing in eukaryotic organisms.
The team used SageELF, an automated sample prep device that generates 12 contiguous fractions from a single DNA or protein sample. It can be used in NGS workflows to build libraries with multiple insert sizes from the same sample, as well as to preserve precious samples. This Bioanalyzer trace of SageELF fractions is excerpted from the scientific poster: