I bookmarked a paper last week that you might find interesting: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3864310/
Although transcriptional and posttranscriptional events are detected in RNA-Seq data from second-generation sequencing, full-length mRNA isoforms are not captured. On the other hand, third-generation sequencing, which yields much longer reads, has current limitations of lower raw accuracy and throughput. Here, we combine second-generation sequencing and third-generation sequencing with a custom-designed method for isoform identification and quantification to generate a high-confidence isoform dataset for human embryonic stem cells (hESCs). We report 8,084 RefSeq-annotated isoforms detected as full-length and an additional 5,459 isoforms predicted through statistical inference. Over one-third of these are novel isoforms, including 273 RNAs from gene loci that have not previously been identified. Further characterization of the novel loci indicates that a subset is expressed in pluripotent cells but not in diverse fetal and adult tissues; moreover, their reduced expression perturbs the network of pluripotency-associated genes. Results suggest that gene identification, even in well-characterized human cell lines and tissues, is likely far from complete.
Is this DNA or RNA sequencing? What do you want to do? Are you interested in de-novo assembly algorithms?
The Iso-Seq in the title suggests they are asking about RNA as it's a transcriptomics library prep method for PacBio
As Daniel pounted out, it is RNAseq data generated using Ribozero illumina and Isoseq pacbio methods. Thanks Daniel.
I want to utilize the benefits of having both datasets in isoform identification, DEG analysis and studying lncRNA.
Hi Mamta,
I need to perform Illumina Truseq and pacbio seq on the same set of sample. Could you let me know more about the sample preparation method you used.
Thanks Annasha Dutta