SMART-Seq Mouse BCR (with UMIs)

Description

SMART-Seq Mouse BCR with UMIs enables users to analyze B-cell receptor BCR repertoires from 10 ng1 mug high-integrity RIN 7 bulk total RNA samples from spleen, bone marrow, or peripheral blood mononuclear cells PBMCs or RNA extracted from mouse whole blood RNA input 100 ng or lymph nodes. The kit can be used to generate data for both heavy IgG, IgM, IgA, IgD, and IgE and light chains IgK, IgL of mouse B-cell receptors. 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 Schematic of technology and workflow for SMART-Seq Muman 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 mBCR PCR1 Universal Forward primer. Following reverse transcription, semi-nested PCR is performed to amplify BCR cDNAs. In PCR1, the mBCR PCR1 Universal Forward primer anneals to the complementary sequence carried by the SMART UMI Oligo light green , incorporating Illumina Read2 sequence dark green . The mBCR 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 mBCR LC light chain or mBCR HC heavy chain PCR2 Reverse primers anneal to sequences in BCR constant regions that are internal to the sequences bound by the mBCR PCR1 Reverse primer and add the Illumina Read 1 sequence light purple . In the same PCR2 reaction, Unique Dual Index Kit primers anneal to sequence added by mBCR PCR1 Universal Forward primer or the mBCR 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 Panel A.Detection of all BCR isotypes for both heavy chain IgA/D/G/M, except IgE due to its very low expression in normal mouse samples and light chain IgK/L by SMART-Seq Mouse BCR with UMIs in 10, 100, and 1,000 ng input of mouse spleen RNA. Comparison across various RNA inputs shows a consistent increase in clonotype counts with higher input.Panel B.Sensitivity of SMART-Seq Mouse BCR with UMIs ensures detection of more heavy-chain and light-chain clonotypes with more input RNA. Bar graphs illustrate both heavy-chain and light-chain clonotype counts consistently increase with a higher amount of input RNA. 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 NucleoMag NGS Clean-up and Size Select employs scalable, automation-friendly magnetic bead technology to enable efficient cleanup of DNA or RNA fragments. The flexible protocol enables processing of 50150 mul input volumes and allows for enrichment of fragments ranging in size from 150800 bp. NucleoMag NGS Clean-up and Size Select consists of paramagnetic beads suspended in binding buffer that selectively bind DNA and RNA fragments based on the volume ratio of bead suspension to sample. After magnetic separation and removal of supernatant, the beads are washed with ethanol followed by a short drying step. Following elution in low-salt buffer or water, the highly purified DNA is free of contaminants such as nucleotides, primers, adapters, adapter dimers, enzymes, buffer additives, or salts, and can be used directly in downstream applications. NucleoMag NGS Clean-up and Size Select provides comparable performance and employs the same dilution ratios for size selection as magnetic bead-based solutions from other providers, such that it can be incorporated in existing workflows without the need for protocol adjustment or optimization. Cat. # 744970.5 consists of a bottle containing 5 ml of bead suspension. Back Back Back Back Back Back NucleoMag NGS Clean-up and Size Select employs scalable, automation-friendly magnetic bead technology to enable efficient cleanup of DNA or RNA fragments. The flexible protocol enables processing of 50150 mul input volumes and allows for enrichment of fragments ranging in size from 150800 bp. NucleoMag NGS Clean-up and Size Select consists of paramagnetic beads suspended in binding buffer that selectively bind DNA and RNA fragments based on the volume ratio of bead suspension to sample. After magnetic separation and removal of supernatant, the beads are washed with ethanol followed by a short drying step. Following elution in low-salt buffer or water, the highly purified DNA is free of contaminants such as nucleotides, primers, adapters, adapter dimers, enzymes, buffer additives, or salts, and can be used directly in downstream applications. NucleoMag NGS Clean-up and Size Select provides comparable performance and employs the same dilution ratios for size selection as magnetic bead-based solutions from other providers, such that it can be incorporated in existing workflows without the need for protocol adjustment or optimization. Cat. # 744970.50 consists of a bottle containing 50 ml of bead suspension. Back Back Back Back Back Back NucleoMag NGS Clean-up and Size Select employs scalable, automation-friendly magnetic bead technology to enable efficient cleanup of DNA or RNA fragments. The flexible protocol enables processing of 50150 mul input volumes and allows for enrichment of fragments ranging in size from 150800 bp. NucleoMag NGS Clean-up and Size Select consists of paramagnetic beads suspended in binding buffer that selectively bind DNA and RNA fragments based on the volume ratio of bead suspension to sample. After magnetic separation and removal of supernatant, the beads are washed with ethanol followed by a short drying step. Following elution in low-salt buffer or water, the highly purified DNA is free of contaminants such as nucleotides, primers, adapters, adapter dimers, enzymes, buffer additives, or salts, and can be used directly in downstream applications. NucleoMag NGS Clean-up and Size Select provides comparable performance and employs the same dilution ratios for size selection as magnetic bead-based solutions from other providers, such that it can be incorporated in existing workflows without the need for protocol adjustment or optimization. Cat. # 744970.500 consists of a bottle containing 500 ml of bead suspension. Back Back Back Back Back Back