Takara SMART-Seq Total RNA Mid Input, 24 Rxns

SMART-Seq Total RNA Mid Input includes the components needed to generate indexed cDNA libraries suitable for next-generation sequencing (NGS) on any Illumina platform, starting from as little as 100 pg of poly(A)-purified or ribosomal RNA-depleted RNA. The directionality of the template-switching reaction preserves the strand orientation of the RNA, making it possible to obtain strand-specific sequencing data from the synthesized cDNA. Indexes are added using a unique dual index kit. The quantity of reagents in this size is sufficient for up to 24 reactions.

cDNA libraries produced using the SMARTer Stranded RNA-Seq Kit show comparable yields and purity irrespective of input RNA concentrations.Agilent 2100 Bioanalyzer gel-like image using a High Sensitivity DNA chip with cDNA libraries produced from between 100 ng–100 pg human brain poly(A)+RNA with the number of PCR cycles indicated. cDNA libraries were produced by amplification with SeqAmp DNA Polymerase (included in the SMARTer Stranded RNA-Seq Kit). Data is shown for the SMARTer Stranded RNA-Seq Kit. SMART-Seq Total RNA Mid Input is an equivalent replacement.

The SMARTer Stranded RNA-Seq Kit provides high reproducibility and sensitivity over a 1,000-fold range of input RNA levels.Scatter plots of expression (FPKM) for cDNA libraries prepared from 100 ng and 100 pg of human brain poly(A)+RNA show a high correlation, suggesting consistency across input levels. Axes are plotted on a log10scale. Insets indicate the Pearson coefficient of correlation between replicates (R). Data is shown for the SMARTer Stranded RNA-Seq Kit. SMART-Seq Total RNA Mid Input is an equivalent replacement.

Data from technical replicates of each library indicate that the SMARTer Stranded RNA-Seq Kit delivers very consistent results across a wide range of input RNA concentrations.Comparisons of pairs of cDNA library replicas created from 100 ng and 100 pg of input RNA show high reproducibility across a wide range of input RNA. Axes are plotted on a log10scale. Insets indicate the Pearson coefficient of correlation between replicates (R). Data is shown for the SMARTer Stranded RNA-Seq Kit. SMART-Seq Total RNA Mid Input is an equivalent replacement.

The SMARTer Stranded RNA-Seq Kit maintains high accuracy even with low input volume.~15,000 genes were detected using strand-specific alignments, even with only 100 pg poly(A)+input RNA. Data is shown for the SMARTer Stranded RNA-Seq Kit. SMART-Seq Total RNA Mid Input is an equivalent replacement.

High reproducibility with the SMARTer Stranded RNA-Seq Kit, confirmed by ERCC analysis.Reads mapping to the ERCC data set from the previous experiments were pooled together and the FPKM was plotted against relative transcript abundance. The complete set of 92 ERCC transcripts was identified, with a slope of 0.934 and R2of 0.9725. Axes are plotted on a log2scale. Data is shown for the SMARTer Stranded RNA-Seq Kit. SMART-Seq Total RNA Mid Input is an equivalent replacement.

Indexed cDNA libraries were prepared according to the SMARTer Stranded RNA-Seq Kit protocol using twelve indices, and sequenced on an Illumina HiSeq® Platform with 2 x 100 bp paired end reads.Short, overlapping reads originating from different strands of the genomic DNA were distinguished from each other, enabling quantitative expression analysis and accurate genome annotation. 99% of RNA-Seq reads mapped to the correct strand. Data is shown for the SMARTer Stranded RNA-Seq Kit. SMART-Seq Total RNA Mid Input is an equivalent replacement.

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. Data is shown for the SMARTer Stranded RNA-Seq Kit. SMART-Seq Total RNA Mid Input is an equivalent replacement.

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 (Takara Bio) 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 (Takara Bio), while retaining more noncoding reads.

Required Products

License Statement

RiboGone - Mammalian allows for the specific removal of rRNA sequences (5S, 5.8S, 18S,and28S), as well as mitochondrial rRNA sequences (12S), from human, mouse, or rat total RNA. This kit is designed for use with limited sample amounts (10–100 ng of total RNA), and works with high- or low-quality RNA. Samples processed using the RiboGone kit are ready for cDNA synthesis with random primers.

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 (Takara Bio) 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 (Takara Bio), while retaining more noncoding reads.

The Unique Dual Index (UDI) Kit (1–96) 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 Bio’s RNA-seq and DNA-seq kits.

The Unique Dual Index (UDI) Kit (97–192) 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 Bio’s RNA-seq and DNA-seq kits.

The Unique Dual Index (UDI) Kit (193–288) 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 Bio’s RNA-seq and DNA-seq kits.

The Unique Dual Index (UDI) Kit (289–384) 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 Bio’s RNA-seq and DNA-seq kits.

The Unique Dual Index (UDI) Kit (1–24) contains 24 pairs of unique dual-indexed PCR primers, which are a subset of the Unique Dual Index Kit (1–96), Cat. No. 634752. They can be used to generate up to 24 Illumina-compatible sequencing libraries and are recommended with several of Takara Bio’s RNA-seq and DNA-seq kits.

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 50–150 µl input volumes and allows for enrichment of fragments ranging in size from 150–800 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.

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 50–150 µl input volumes and allows for enrichment of fragments ranging in size from 150–800 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.

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 50–150 µl input volumes and allows for enrichment of fragments ranging in size from 150–800 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.