SMARTer Stranded Total RNA Sample Prep Kit - Low Input Mammalian
SMARTer Stranded Total RNA Sample Prep Kit - Low Input Mammalian
Brand: Takara Bio.
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Stranded Total RNA Sample Prep Kit - Low Input Mammalian
Strand-specific Illumina sequencing libraries from low RNA inputs of any quality
The SMARTer Stranded Total RNA Sample Prep Kit - Low Input Mammalian provides a complete solution for sensitive and streamlined total RNA analysis. The kit is designed to work with a wide range of input amounts (10–100 ng) of total RNA of any quality. A variety of human, rat, and mouse samples can be used, including total RNA from FFPE material, tissue samples, etc. The kit provides whole transcriptome analysis with accurate measurement of strand orientation, unbiased coverage, high transcript mapping, and gene counts.
This low-input kit contains the Illumina Indexing Primer Set, and the entire protocol—from rRNA depletion through the creation of Illumina-specific, indexed RNA-seq libraries—takes only about 5 hours.
Overview
- Strand information—Identify each transcript’s strand of origin with >99% accuracy.
- Fast, streamlined protocol—Go from start to finish in ~5 hours.
- Variable input size—Use 10–100 ng of total RNA from human, mouse, or rat.
- Versatility—Get reliable data across replicates. See high-quality, reproducible data from total RNA with a RIN (RNA Integrity Number) from 3–10.
- Seamless integration with Illumina sequencing—Generate 24 uniquely indexed Illumina-ready libraries.
Applications
- Robust NGS library construction that retains strand information
- Use for RNA-seq on all Illumina platforms
- Captures coding and noncoding information from total mammalian RNA of any quality, including RNA obtained from FFPE samples
Distinguishing overlapping and antisense transcripts with the SMARTer Stranded RNA-Seq Kit
Distinguishing overlapping and antisense transcripts with the SMARTer Stranded RNA-Seq Kit. Panel A. RNA-seq reads from the Human Brain Poly A+ RNA cDNA library were mapped against the human genome. The SMARTer Stranded method allowed assignment of sequencing reads to the correct gene in the case of overlapping PHC1 and M6PR transcripts. Panel B. Strand-specific coverage of the CDR1 locus. Nearly all reads are antisense to the annotated transcript, a finding independently reported elsewhere (Hansen, T. B. et al. (2011) EMBO J. 30(21):4414–4422). Panel C. Comparison of CDR1 gene counts obtained using either a strand-agnostic or strand-aware method.
RiboGone - Mammalian removes rRNA efficiently from intact and degraded RNA, while retaining noncoding transcripts for analysis
RiboGone - Mammalian removes rRNA efficiently from intact and degraded RNA, while retaining noncoding transcripts for analysis. RNA-seq libraries were generated from Human Brain Total RNA (Clontech) or Breast Cancer FFPE RNA (Cureline, extracted using a NucleoSpin totalRNA FFPE kit) and treated with the indicated rRNA removal method. Reads were mapped to the hg19 genome and read distributions were determined using Picard RNA-Seq Metrics. Libraries generated from RiboGone-treated RNA had comparably low rRNA reads to oligo(dT)-enriched RNA (Clontech), while retaining more noncoding reads.
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Distinguishing overlapping and antisense transcripts with the SMARTer Stranded RNA-Seq Kit
Distinguishing overlapping and antisense transcripts with the SMARTer Stranded RNA-Seq Kit. Panel A. RNA-seq reads from the Human Brain Poly A+ RNA cDNA library were mapped against the human genome. The SMARTer Stranded method allowed assignment of sequencing reads to the correct gene in the case of overlapping PHC1 and M6PR transcripts. Panel B. Strand-specific coverage of the CDR1 locus. Nearly all reads are antisense to the annotated transcript, a finding independently reported elsewhere (Hansen, T. B. et al. (2011) EMBO J. 30(21):4414–4422). Panel C. Comparison of CDR1 gene counts obtained using either a strand-agnostic or strand-aware method.
RiboGone - Mammalian removes rRNA efficiently from intact and degraded RNA, while retaining noncoding transcripts for analysis
RiboGone - Mammalian removes rRNA efficiently from intact and degraded RNA, while retaining noncoding transcripts for analysis. RNA-seq libraries were generated from Human Brain Total RNA (Clontech) or Breast Cancer FFPE RNA (Cureline, extracted using a NucleoSpin totalRNA FFPE kit) and treated with the indicated rRNA removal method. Reads were mapped to the hg19 genome and read distributions were determined using Picard RNA-Seq Metrics. Libraries generated from RiboGone-treated RNA had comparably low rRNA reads to oligo(dT)-enriched RNA (Clontech), while retaining more noncoding reads.
