Thermostable MMLV-RT

55C MMLV-RT

GC-rich Regions

Robust thermostable reverse transcriptase ideal for ambient temperature stable molecular assays.

Reverse transcriptase activity up to 60°C

Ideal for RNA with high secondary structure such as viral genomes

Sensitive detection of low copy number RNA targets

Helices

Formulation compatible with lyophilization and air-drying applications

Challenges with RNA Secondary Structures

Ideal for developing fast, highly reproducible RT-qPCR assays

Product Information

PRODUCT

CAT NO.

VOLUME

REACTIONS

50 µL

10,000 Units

55C MMLV-RT Thermostable Moloney Murine Leukemia Virus (concentration 200 U/µL)

200 µL

40,000 Units

MDX117

1 mL

200,000 Units

10 mL

2,000,000 Units

Performance Data

High Thermostability

Traditional Moloney Murine Leukemia Virus Reverse Transcriptase (MMLV-RT) is not thermostable and can only maintain its enzymatic activity at relatively low temperatures (up to 50°C). However for cDNA synthesis, a higher reaction temperature is desirable as it reduces RNA secondary structures which can inhibit reverse transcription and it minimizes nonspecific primer binding. Meridian has developed a new 55C MMLV-RT that has higher thermal stability and reduced RNase H activity. The enzyme can be used to synthesize first-strand cDNA at temperatures up to 60°C which improves the cDNA yield from difficult RNA targets that require higher temperature to denature strong secondary structures.

Comparison of thermostability of 55C MMLV-RT vs other RTases

Performance of Meridian’s 55C MMLV-RT (red) , standard MMLV-RT (grey) and a thermostable reverse transcriptase from supplier X (black) after pre-incubation at 40°C to 70°C for 10 mins, in a multiplex one-step RT-qPCR assay. ∆ ct values were calculated against the ct value produced by the same enzyme stored at -20 °C. The data illustrates the increased thermal stability of 55C MMLV-RT when compared to other MMLV-RT enzymes and its ability to efficiently synthesize cDNA at temperatures up to 60°C.

MMLV-RT | Supplier X | 55C MMLV-RT

www.MeridianLifeScience.com

Higher Enzyme Efficiency and Sensitivity

55C MMLV-RT is designed for greater efficiency of the reverse transcription reaction, enabling a lower limit of detection (LOD) with higher sensitivity in one-step RT-qPCR assays.

B2MG

Rps18L

55C MMLV-RT | Supplier X

The sensitivity of 55C MMLV-RT (red) was compared to supplier X (black) in a multiplex one-step RT-qPCR assay using a 10-fold serial dilution of mammalian total RNA. The results demonstrate that 55C MMLV-RT has higher performance with better sensitivity and end-fluorescence.

Compatible with Lyophilization and Air-drying Applications

55C MMLV-RT proprietary formulation allows for incorporation into assays designed for subsequent lyophilization or air-drying.

Flu A

MERS-CoV

RSV

Dried Mix | Wet Mix

55C MMLV-RT was added to an air-dryable RT-qPCR Mix and dried down in a fan assisted oven (red) . After rehydration, the mix was tested against freshly prepared (wet) version of this mix (blue) , in a triplex one-step RT-qPCR assay on respiratory RNA virus targets (Influenza A, MERS-CoV and RSV). The results demonstrate that 55C MMLV-RT retains the same activity level after air-drying even in challenging conditions such as multiplex RT-qPCR reactions.

Ordering information: USA 5171 Wilfong Road Memphis, Tennessee 38134 Fax: +1 901-333-8223 Toll Free: +1 800 327 6299

Email: info@meridianlifescience.com Orders: orders@meridianlifescience.com www.MeridianLifeScience.com

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