RNA-Seq of single cells has been limited by biases in transcript coverage and unknown technical variability. Ramsköld et al. describe a protocol to reproducibly recover full-length transcripts and use it to quantitatively analyze splice isoforms in si
从建库测序原理上区分,主要有两大技术方案:一是以 drop-seq 为代表的3‘/ 5’端测序,这种方法虽然只能测到转录本一端大约600nt以内的序列,但是在大多数情况下,用部分序列代表整条分子用于 mRNA 的定量是可行的;第二种技术是以 smart-seq 技术为代表的...
Here we describe a robust mRNA-Seq protocol (Smart-Seq) that is applicable down to single cell levels. Compared with existing methods, Smart-Seq has improved read coverage across transcripts, which enhances detailed analyses of alternative transcript isoforms and identification of s...
RNA-Seq of single cells has been limited by biases in transcript coverage and unknown technical variability. Ramsköld et al. describe a protocol to reproducibly recover full-length transcripts and use it to quantitatively analyze splice isoforms in si
NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript Full-Length mRNA-Seq from single cell levels of RNA and individual circulating tumor cells Daniel Ramsk?ld1,2,*, Shujun Luo3,*, Yu-Chieh Wang4, Robin Li3, Qiaolin Deng1, Omid R. Faridani1, Gregory A. Daniels...
full-length mRNA sequencing (Additional file1: Figure S7, 8, 10, and 11). In addition, we carried out a single-molecule RNA in situ fluorescence (smRNA FISH) co-localization approach [21,22] to examine the alternative splicing events that were identified by full-length RNA-seq and ...
Emerging methods for the accurate quantification of gene expression in individual cells hold promise for revealing the extent, function and origins of cell-to-cell variability. Different high-throughput methods for single-cell RNA-seq have been introduced that vary in coverage, sensitivity and multiplex...
“Spliceosome”, “Carbon metabolism”, “mRNA surveillance pathway” and “Biosynthesis of amino acids” (Fig.2A, C). All the differential AS genes sets were enriched in the spliceosome. Glyoxylate and dicarboxylate metabolism was found to be a unique pathway in R, while Carbon metabolism was ...
transcriptomegenomeassemblyreferenceseqlength naturebiotechnologyadvanceonlinepublication1 1 BroadInstitutofMassachusttsInstitutofTchnologyandHarvard,Cambridg,Massachustts,USA. 2 SchoolofComputrScinc,HbrwUnivrsity,Jrusalm, Isral. 3 DpartmntofBiology,MassachusttsInstitutofTchnology,Cambridg,Massachustts,USA. 4 Dpa...
Massively parallel sequencing of cDNA has enabled deep and efficient probing of transcriptomes. Current approaches for transcript reconstruction from such data often rely on aligning reads to a reference genome, and are thus unsuitable for samples with a partial or missing reference genome. Here we pr...