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Related Experiment Video

Updated: Jan 19, 2026

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Comparative evaluation of RNA-Seq library preparation methods for strand-specificity and low input.

Dimitra Sarantopoulou1, Soon Yew Tang2, Emanuela Ricciotti2

  • 1Institute for Translational Medicine and Therapeutics, University of Pennsylvania, Philadelphia, PA, USA.

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|September 19, 2019
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Summary

The Takara Bio Pico kit enables strand-specific RNA sequencing with low input material, unlike other kits. It provides comparable pathway analysis results to higher-input methods, making it a versatile option for gene expression studies.

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Area of Science:

  • Molecular Biology
  • Genomics
  • Bioinformatics

Background:

  • Library preparation is crucial for RNA sequencing.
  • Strand specificity and low input material are desirable but often mutually exclusive in RNA library prep kits.
  • Existing kits like Illumina TruSeq and Takara Bio SMART-Seq v4 have limitations regarding input material or strand specificity.

Purpose of the Study:

  • To comparatively evaluate the Takara Bio SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian (Pico) against established kits.
  • To assess the performance of the Pico kit in differential gene expression analysis using minute starting material while maintaining strand specificity.
  • To determine if the Pico kit offers a viable alternative for researchers needing both strand specificity and low input RNA sequencing.

Main Methods:

  • Comparative analysis of three RNA library preparation kits: Illumina TruSeq stranded mRNA Sample Preparation kit (TruSeq), Takara Bio SMART-Seq v4 Ultra Low Input RNA kit (V4), and Takara Bio SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian (Pico).
  • Preparation and sequencing of RNA samples from two experimental conditions using each of the three kits.
  • Differential gene expression analysis and pathway enrichment analysis to compare kit performance.

Main Results:

  • The Pico kit identified 55% fewer differentially expressed genes compared to the TruSeq kit.
  • Despite differences in gene-level expression, comparable functional results were observed through enriched pathway analysis across all tested kits.
  • The Pico kit successfully combined strand specificity with ultra-low input material, a feature lacking in the other evaluated kits.

Conclusions:

  • The Takara Bio Pico kit is a suitable option for RNA sequencing, effectively reproducing pathway analysis results comparable to other kits.
  • It offers a unique advantage by enabling both strand-specific sequencing and the use of minute starting material.
  • Researchers can confidently use the Pico kit for differential gene expression studies, particularly when input RNA is limited.