Series | GSE38495 |
Title | Full-length mRNA-Seq from single-cell levels of RNA and individual circulating tumor cells |
Year | 2012 |
Country | Sweden |
Article | Sandberg R,Schroth GP,Laurent LC,Loring JF,Khrebtukova I,Daniels GA,Faridani OR,Deng Q,Li R,Wang YC,Luo S,Ramsköld D.Full-length mRNA-Seq from single-cell levels of RNA and individual circulating tumor cells.Nature biotechnology.2012 Aug |
PMID | 22820318 |
Bio Project | BioProject: http://www.ncbi.nlm.nih.gov/bioproject/PRJNA170814 |
Sra | SRA: http://www.ncbi.nlm.nih.gov/sra?term=SRP014428 |
Overall Desgin | We generated RNA-Seq libraries for dilution series of MAQC reference RNA and mouse brain RNA to assess technical reproducibility, and for a variety of individual cells including putative circulating tumour cells. |
Summary | We assessed Smart-Seq, a new single-cell RNA-Seq library preparation method, on a variety of mouse and human RNA samples or cells. |
Experimental Protocol | cDNA was generated as described in the manual for SMARTer Ultra Low RNA Kit for Illumina sequencing marketed by Clontech. Then, a sequencing library was generated as described in Illumina's Ultra Low Input mRNA-Seq Guide. cDNA was generated as described in the manual for SMARTer Ultra Low RNA Kit for Illumina sequencing marketed by Clontech. Then, a sequencing library was generated using the Epicentre Nextera sample preparation kit, with 5 min tagmentation at 55C with 0.25 ul transposase and 4 ul 5x HMW Nextera reaction buffer, then 35 ul of PB, then 88 ul SPRI XP beads.
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Data processing | Alignment by Bowtie (version 0.12.7, option –best) to genome and exon-exon junctions.; Filtering for uniquely mapping reads (removal of reads aligning with same number of mismatches to several coordinates).; Conversion by samtools to bam format.; Conversion by bedtools to bed format.; Generation of RPKM (reads per kilobase and million mapped reads) values by rpkmforgenes.py available at http://sandberg.cmb.ki.se/rnaseq/.; Genome_build: mm9 for mouse; hg19 for human.; Supplementary_files_format_and_content: Bed12 format with uniquely aligned reads.; Supplementary_files_format_and_content: Tab-delimited plain text format with gene expression values in RPKM.
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Platform | GPL10999;GPL11002;GPL13112;GPL15520 |
Public On | Public on Jul 22 2012 |
Differential expression gene List between two groups.