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...
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
Full-length mRNA-Seq from single-cell levels of RNA and individual circulating tumor cells.AnimalsHumansMiceMelanomaRNA, MessengerOligonucleotide Array Sequence AnalysisCluster AnalysisSensitivity and SpecificityGene Expression ProfilingSequence Analysis, RNA...
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
full-length mRNA sequencing data can be reliably used for locus-specific quantification in MCF-7 cells. To further establish the accuracy of the transcriptional start sites (TSSs) and polyadenylation sites (PASs) obtained with the Iso-Seq method, we performed a transcriptome-wide comparison of 5...
To enable gene and mRNA isoform expression analyses in single cells, a novel full-transcriptome mRNA-Seq protocol (Smart-Seq) was developed. Smart-Seq makes use of SMART? template switching technology for the generation of full-length cDNAs and only 12 to 18 cycles of PCR following the ...
seq for transcriptome analysis from single cells, and we subsequently optimized the method for improved sensitivity, accuracy and full-length coverage across transcripts. Here we present a detailed protocol for Smart-seq2 that allows the generation of full-length cDNA and sequencing libraries by using...
Here, we present full-length poly(A) and mRNA sequencing (FLAM-seq), a rapid and simple method for high-quality sequencing of entire mRNAs. We report a complementary DNA library preparation method coupled to single-molecule sequencing to perform FLAM-seq. Using human cell lines, brain organoid...
transcriptomegenomeassemblyreferenceseqlength naturebiotechnologyadvanceonlinepublication1 1 BroadInstitutofMassachusttsInstitutofTchnologyandHarvard,Cambridg,Massachustts,USA. 2 SchoolofComputrScinc,HbrwUnivrsity,Jrusalm, Isral. 3 DpartmntofBiology,MassachusttsInstitutofTchnology,Cambridg,Massachustts,USA. 4 Dpa...
and are thus unsuitable for samples with a partial or missing reference genome. Here we present the Trinity method forde novoassembly of full-length transcripts and evaluate it on samples from fission yeast, mouse and whitefly, whose reference genome is not yet available. By efficiently constructing...