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TransNGS Stranded RNA-Seq Library Prep Kit for Illumina – KP301

KP301 Stranded RNA Library Prep Workflow and time Flow

$457.00 – $5,171.00 CAD

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Stranded RNA Seq Library Prep Kit for Illumina

TransNGS Stranded RNA-Seq Library Prep Kit for Illumina is designed to prepare Stranded RNA libraries compatible with the Illumina® sequencers. Differently from conventional stranded RNA seq library prep kits, the TransNGS Stranded RNA seq kit generates cDNA using random hexamer with a 3’-Adaptor. Subsequently, the 5’-Adaptor is ligated at the 3’-end of the first-strand cDNA, bypassing the need for 2nd strand synthesis and digestion. Then, end-repair and dA-tailing are performed. As a result, only information from the 1st strand cDNA is preserved. This kit is tested and promised to provide stable and reproducible cDNA libraries, by using either total RNA samples, rRNA-depleted samples, mRNA purified samples or lncRNA samples.

Highlights

  • User-friendly and fast with minimal hands-on time
  • High efficiency with minimal amount of input DNA
  • Operation is simple and fast

Applications

  • Gene expression analysis
  • Single–nucleotide variation (SNV) discovery
  • Splice junction and gene fusion identification
  • Whole transcriptome analysis
  • Characterization of non-polyadenylated RNAs, including non-coding and immature RNAs

Kit Contents

Components KP301-01 (6rxns) KP301-02 (24rxns) KP301-03 (96rxns)
2×RNA-Seq RT Reaction Mix 60 μl 240 μl 4×240 μl
RNA-Seq RT Enzymes Mix 6 μl 24 μl 4×24 μl
RNA Digestion Mix 6 μl 24 μl 4×24 μl
cDNA Binding Buffer 180 μl 720 μl 4×720 μl
5′-Adaptor 6 μl 24 μl 96 μl
2×RNA-Seq Ligation Mix 150 μl 600 μl 4×600 μl
RNA-Seq Ligase 30 μl 120 μl 480 μl
RNA Ligation Stop Buffer 200 μl 800 μl 4×800 μl
TransNGSTM Library Amplification SuperMix 150 μl 600 μl 4×600 μl
Library Elution Buffer (LEB) 1.2 ml 5 ml 4×5 ml
RNase-free Water 1 ml 1 ml 5 ml

Cat. No. KP301

Storage: at -20°C for one year

RNA Input Recommendations

Starting Materials: 10 ng – 1 μg total RNA treated with rRNA depletion, 0.1 μg – 4 μg total RNA treated with mRNA purification and lncRNA.

Requirements: high quality RNA, which is recommended to use an Agilent 2100 Bioanalyzer to analyze the total RNA. The RIN (RNA integrity number) value should be ≥ 7.0. Using degraded total RNA for library construction will lead to in 3’ bias in RNA-seq. The ratio of OD260/OD280 should be between 1.8 and 2.1.

Reagents not included in the kit

  1. 80% ethanol (freshly prepared);
  2. DNA size selection beads like MagicPure Size Selection DNA Beads (TransGen, EC401);
  3. Index Primers : TransNGS Single Index Primers Kit for Illumina® (TransGen, KI201-KI221) or Dual Index Primers Kit for Illumina® (TransGen, KI231).

Workflow for Stranded RNA library prep

KP301 Stranded RNA Library Prep Workflow and time Flow

Manual & Datasheet

KP301 – TransNGS Stranded RNA-Seq Library Prep Kit for Illumina – 2018-11-07

Additional information

Format

6 rx, 24 rx, 96 rx
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