SMARTer Target RNA Capture for Illumina
SMARTer Target RNA Capture for Illumina
Brand: Takara Bio.
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SMARTer Target RNA Capture for Illumina
SMARTer Target RNA Capture for Illumina
SMARTer Target RNA Capture for Illumina is a complete system is designed to generate cDNA from enriched total RNA. Targeted RNA-seq can overcome several challenges inherent in whole transcriptome amplification (WTA) RNA-seq experiments, improving sequence coverage and sensitivity of detection of transcripts of interest that may be present in low amounts. Targeted RNA-seq approaches are also advantageous in saving costs and simplifying analysis. Enrichment of transcripts of interest prior to cDNA synthesis enables the capture of information about transcripts that could otherwise be missed or require a much greater number of sequencing reads to be detected, including chimeric gene fusions, transcript isoforms, and splice variants. By improving sensitivity, the power of analysis is also improved, as seen in a reduction in the signal-to-noise ratio.
In the SMARTer Target RNA Capture for Illumina workflow, total RNA is hybridized with user designed biotinylated DNA probes in less than four hours and then purified prior to capture on Capture Beads—magnetic particles coated with streptavidin. The Capture Beads have been selected because they exhibit especially low adsorption of protein and nucleic acids, and do not interfere with downstream reactions such as PCR or reverse transcription, making it possible to perform cDNA synthesis on the targeted samples while they remain attached to the beads. Capture Beads are provided as part of the SMARTer Target RNA Capture for Illumina kit, and are also sold separately (Cat. # 635039).
Overview
- Inputs as low as 10 ng of purified total RNA
- Streamlined workflow for specific enrichment of transcripts of interest
- Generate Illumina-ready, full-length sequencing libraries from the enriched DNA
- Ligation-independent sequencing adapter addition for library preparation
Applications
- Sequencing library preparation for targeted RNA-seq from purified total RNA
- Illumina-specific NGS sequencing library production
- Detection of rare gene fusion events at low sequencing depth
SMARTer Target RNA Capture for Illumina protocol overview
SMARTer Target RNA Capture for Illumina protocol overview. The protocol through validation is completed over two days, with just 2.5–3 hours of hands-on time and without the need for additional rRNA removal methods or kits.
Example electropherogram results from Agilent 2100 Bioanalyzer
Example electropherogram results from Agilent 2100 Bioanalyzer. Unless indicated, 10 ng of Control Total RNA was subjected to hybridization with HPRT1 Control Probes, capture with Capture Beads, SMART-Seq v4 cDNA synthesis, and amplification for 20 cycles as described in the protocol. Panel A shows a single distinct peak between 1,400–1,650 bp. Panels B and C show no product in the negative controls (no RNA and no probes, respectively). Panel D shows the cDNA profile resulting from 10 ng of Control Total RNA hybridized to probes for 10 different genes, following amplification for 17 cycles.
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SMARTer Target RNA Capture for Illumina protocol overview
SMARTer Target RNA Capture for Illumina protocol overview. The protocol through validation is completed over two days, with just 2.5–3 hours of hands-on time and without the need for additional rRNA removal methods or kits.
Example electropherogram results from Agilent 2100 Bioanalyzer
Example electropherogram results from Agilent 2100 Bioanalyzer. Unless indicated, 10 ng of Control Total RNA was subjected to hybridization with HPRT1 Control Probes, capture with Capture Beads, SMART-Seq v4 cDNA synthesis, and amplification for 20 cycles as described in the protocol. Panel A shows a single distinct peak between 1,400–1,650 bp. Panels B and C show no product in the negative controls (no RNA and no probes, respectively). Panel D shows the cDNA profile resulting from 10 ng of Control Total RNA hybridized to probes for 10 different genes, following amplification for 17 cycles.
