MeloScript Reverse Transcriptase

Product Descriptions

MeloScript Reverse Transcriptase is an engineered MMLV RT-based reverse transcriptase. MeloScript RT can perform reverse transcription at high temperature (optimal at 55℃ and can go up to 65℃）to transcribe through RNA secondary structure and high GC region. In addition, Meloscript RT shows superior sensitivity, high inhibitor resistance and specificity.

Specifications & Components

Applications

• Synthesis of the first-strand cDNA.

• One-step RT-qPCR.

Unit Definition

One unit is defined as the amount of enzyme required to incorporate 1nmol of dTTP into an acid-insoluble form in 10 minutes at 37°C using poly (rA)-oligo (dT) as template primer.

Highlights of MeloScript

• Superior universal performance with a variety of target amplicons

• Thermostability with reaction temperature up to 65℃

• Strong inhibitor resistance

• High sensitivity in multiplex PCR/qPCR reactions

• High quality enzyme with consistent performance and minimal batch-to-batch variation

Key Performance Data

MeloScript Target Adaptability Assessment

The performance of various reverse transcriptases were evaluated with the target adaptability assessment assay, where each amplicon represented by each triangle (24 amplicons in total) was scored based on PCR efficiency, dynamic range, target specificity, and accuracy with PCR efficiency on the x-axis and delta Cq (ΔCq) on the y-axis. Only triangles in the dotted box with quality scores >4 are considered as “pass”. MeloScript RT shows superior adaptability with 95% passing rate.

MeloScript RT Inhibitor Tolerance

Various inhibitors were used in RT reaction followed by qPCR in a two-step RT-qPCR reaction. The Ct values were normalized to MeloScript RT. The normalized value lower than 1 indicates less inhibitor resistance and higher than 1 indicates higher inhibitor resistance. MeloScript RT shows overall superior inhibitor resistance property.

Thermostability of MeloScript RT

The thermostability of reverse transcriptase was evaluated in an RNA ladder assay where an RNA ladder (1kb, 2kb and 3kb) was reverse transcribed to cDNA followed by electrophoresis under various reaction temperature. Among all RTs tested, MeloScript RT is the only RT that showed superior thermostability even at 65 °C.