TeloScript Reverse Transcriptase

Description

Teloscript Reverse Transcriptase is a proprietary MMLV mutant exhibiting improved superior processivity and inhibitor resistance in RT reactions. It does not require increased reaction temperature to overcome secondary structures in RNA, thus preserving RNA integrity in complex samples for first-strand cDNA synthesis. In addition, TeloScript RT exhibits superior template switching efficiency, which can be used for single-cell RNA sequencing.

Specifications & Components

Applications

• First-Strand cDNA synthesis

• RNA sequencing

• Single cell RNA sequencing

• 2-step RT-qPCR

Unit Definition

One unit is defined as the amount of enzyme required to incorporate 1 nmol of dTTP into acid-insolublematerial in 10 minutes at 37°C using poly(A)/oligo (dT) as a substrate.

Highlights

• Rapid and highly efficient full-length cDNA synthesis: up to 15 kb in 30 minutes at 42℃.

• Reverse transcription of RNA rich in secondary structure at low temperature: 9 kb HCV RNA template in 15 minutes at 42℃.

• Significantly improved inhibitor resistance.

• Strong template switching activity for single cell RNA seq.

Key Performance Data

TeloScript RT Can Elongate Long or Highly-structured RNA Template

Comparison of elongation activity of TeloScript RT and other commercially available RTs in the RNA ladder assay.  First strand cDNA synthesis activities (A) at 42℃ for 30' using RNA ladder (1, 2, 3, 5, 7, 9, and 15 kb); (B) at 42℃ for 15' using HCV RNA ladder (1, 2, 3, 5, 7, and 9kb, strong secondary structure) as templates.

TeloScript Shows Strong Inhibitor Tolerance

Comparison of inhibitor resistance activity of TeloScript RT and other RTs. First strand cDNA synthesis using RNA ladder as template with different common inhibitors. The reaction was carried out at 42℃ for 30 min.

TeloScript RT Exhibits Strong Template Switching Activity

Performance of TeloScript RT on the template-switching-based cDNA synthesis and amplification step for single-cell or low-input RNA sequencing. The cDNA synthesis and template switching step for low RNA input (10 pg UHR total RNA) was carried out at 42℃ for 90 min by TeloScript RT. Then the first strand cDNA was amplified with 18 PCR cycles and cleaned with beads.  The process with TeloScript RT produced higher yield than the other commercially available RT (n=5) (A), which was confirmed by a representative electropherogram.