SMART-Seq Human BCR (with UMIs)

Description

SMART-Seq Human BCR with UMIs generates oligo dT -primed, full-length mRNA-seq libraries for all BCR heavy IgG/M/D/E/A and light IgK/L chains. The incorporation of unique molecular identifiers UMIs provides greater accuracy for quantitative gene expression analysis across samples while controlling for PCR errors and amplification biases. Up to 384 multiplexed, Illumina-ready sequencing libraries can be obtained using theUnique Dual Index Kits Cat. # 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 Sensitive and reproducible clonotype detection from a broad range of RNA amounts. Human BCR heavy chain IgA/D/E/G/M and light chain IgK/L libraries were generated from 10, 100, and 1,000 ng of total human PBMC RNA using SMART-Seq Human BCR with UMIs . The sequence reads were processed using Cogent NGS Immune Profiler Software. Back Schematic of technology and workflow for SMART-Seq Human BCR with UMIs .Input RNA or cells are oligo-dT primed using the dT Primer dark blue . Following oligo dT-priming, SmartScribe Reverse Transcriptase performs first strand cDNA synthesis on input RNA or cells and adds non-templated nucleotides to the 5 end of each cDNA molecule XXXXX . Upon reaching the 5 end of the RNA template, the SMART UMI Oligo anneals to the non-templated nucleotides XXXXX , incorporating the UMI yellow and partial Illumina adapter light green complementary to the hBCR PCR1 Universal Forward primer. Following reverse transcription, semi-nested PCR is performed to amplify BCR cDNAs. In PCR1, the hBCR PCR1 Universal Forward primer anneals to the complementary sequence carried by the SMART UMI Oligo light green , incorporating the Illumina Read2 sequence dark green . The hBCR PCR1 Reverse primer orange anneals to sequences in the constant regions of BCR cDNAs to amplify the entire BCR V D J region. During the PCR2, the nested hBCR LC light chain or hBCR HC heavy chain PCR2 Reverse primers anneal to sequences in BCR constant regions that are internal to the sequences bound by the hBCR PCR1 Reverse primer and add the Illumina Read 1 sequence light purple . In the same PCR2 reaction, Unique Dual Index Kit primers anneal to the sequence added by hBCR PCR1 Universal Forward primer or the hBCR LC or HC PCR2 Reverse primers to add Illumina P7-i7 and P5-i5 index sequences dark blue . The result is a sequencing-ready library that contains the entire BCR light chain and/or heavy chain variable regions with a small portion of the constant region. Back Back Back Back Back 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