SMART-Seq mRNA LP (with UMIs)

Description

SMART-Seq mRNA LP with UMIs generates oligo dT -primed, full-length mRNA-seq libraries with UMIs, providing greater accuracy for quantitative gene expression analysis across samples, while controlling for PCR errors and amplification biases. The chemistry is optimized for use on ultra-low amounts of total RNA 10 pg100 ng, RIN8 or for direct use on multiple intact cells <1,000 cells . Up to 384 multiplexed, Illumina-ready sequencing libraries can be obtained using theUnique Dual Index kits Cat. Nos. 63475256 . This kit offers an end-to-end solution including cDNA synthesis, library preparation, and data analysis with our free Cogent NGS bioinformatics tools. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back The SMART-Seq chemistry efficiently captures the 5 end of transcripts. UMIs are added during the first strand synthesis. High-quality, Illumina-ready sequencing libraries can be prepared from single cells and ultra-low amounts of RNA. Back The original SMART-Seq mRNA LP kit has an update. The addition of UMIs provides higher accuracy RNA counting. Detect rare transcripts and identify rare cell types with higher confidence. Back The SMART-Seq mRNA LP with UMIs kit workflow goes from cDNA synthesis to library prep. Sequence on an Illumina instrument and use the resulting FASTQ files as input for our free analysis software. Cogent NGS Analysis Pipeline is used for mapping and reporting. Cogent NGS Discovery Software is used for interactive data visualization. Back SMART-Seq mRNA LP with UMIs and SMART-Seq mRNA LP were each used to create sequencing libraries from K562 RNA 10 pg or universal human reference UHR RNA 100 pg, 1 ng, 10 ng, and 100 ng . Gene counts and RNA subtype representation read distribution were similar across the two methods. Back cDNA and sequencing libraries were generated from 10 pg universal human reference UHR RNA using the SMART-Seq mRNA LP with UMIs SSmRNA +UMIs or the SMART-Seq2 SS2 method. The latter were prepared using the Nextera XT DNA Library Prep Kit Illumina . After pooling, sequencing, and downsampling to 1.2 million reads, SSmRNA +UMIs resulted in a higher gene count with similar read distribution. Back Absolute read counts were calculated, and regression analysis was performed for each replicate pair using our free bioinformatic software, Cogent AP. The R2values were consistently higher for SMART-Seq mRNA LP with UMIs than for Smart-seq2. Back 10 pg of input RNA or RNA from a single cell was spiked with ERCC control RNA. Libraries were prepared using the SSmRNA +UMIs method. After sequencing, our free bioinformatic software, CogentAP, was used to plot TPM or CPM against the absolute number of input molecules from the ERCC spike-in. Subsequent regression analysis showed a significant increase in the R2value upon UMI collapse. Back Single peripheral blood mononuclear cells PBMCs were sorted into plates. Sequencing libraries were produced using either SMART-Seq mRNA LP or SMART-Seq mRNA LP with UMIs . Libraries were then pooled, sequenced, and downsampled to 900,000 reads for analysis. The comparability of gene counts read distributions illustrates that data quality is not compromised by including UMIs. Back Back Required Products The Unique Dual Index UDI Kit 196 contains 96 pairs of unique dual-indexed PCR primers that can be used to generate up to 96 Illumina-compatible sequencing libraries and are recommended with several of Takara Bios RNA-seq and DNA-seq kits. Back The Unique Dual Index UDI Kit 97192 contains 96 pairs of unique dual-indexed PCR primers that can be used to generate up to 96 Illumina-compatible sequencing libraries and are recommended with several of Takara Bios RNA-seq and DNA-seq kits. Back The Unique Dual Index UDI Kit 193288 contains 96 pairs of unique dual-indexed PCR primers that can be used to generate up to 96 Illumina-compatible sequencing libraries and are recommended with several of Takara Bios RNA-seq and DNA-seq kits. Back The Unique Dual Index UDI Kit 289384 contains 96 pairs of unique dual-indexed PCR primers that can be used to generate up to 96 Illumina-compatible sequencing libraries and are recommended with several of Takara Bios RNA-seq and DNA-seq kits. Back The Unique Dual Index UDI Kit 124 contains 24 pairs of unique dual-indexed PCR primers, which are a subset of the Unique Dual Index Kit 196 , Cat. No. 634752. They can be used to generate up to 24 Illumina-compatible sequencing libraries and are recommended with several of Takara Bios RNA-seq and DNA-seq kits. Back