描述

Thermo Scientific Maxima Reverse Transcriptase (RT) was developed through in vitro evolution of M-MuLV RT. The enzyme possesses an RNA-dependent and DNA-dependent polymerase activity as well as RNase H activity. The engineered enzyme features dramatically improved thermostability and robustness, and increased synthesis rates compared to wild type M-MuLV RT.

Maxima Reverse Transcriptase is capable of reproducible cDNA synthesis from a wide range of input total RNA amounts (from 1 pg to 5 µg) at elevated temperatures (50 to 65°C), which makes this enzyme an ideal tool for two step RT-qPCR (see supporting data).

Due to its high thermostability, Maxima enzyme maintains full activity during the entire reverse transcription reaction, generates the highest yields of cDNA, and is able to synthesize even very long RNA transcripts up to 20 kb. The reaction temperature can be increased up to 65°C for efficient transcription of RNA regions with a high secondary structure or to improve specificity using gene specific primers. Due to its increased synthesis rate, the reverse transcriptase reaction can be completed in 15 to 30 min.

Highlights

• High yields of full-length cDNA up to 20 kb
• Active up to 65°C
• Thermostable—90% active after incubation at 50°C for 60 minutes
• High sensitivity—reproducible cDNA synthesis from a wide range of total RNA quantities (1 pg to 5 µg)
• Efficient—complete cDNA synthesis in 15 to 30 minutes
• Incorporates modified nucleotides

Applications

• Two step RT-PCR
• Two step RT-qPCR
• First strand cDNA synthesis
• Construction of full length cDNA libraries
• DNA labeling
• Primer extension

Also available: Maxima First Strand cDNA Synthesis Kit for RT-qPCR

High thermostability of Maxima Reverse Transcriptase at 50°C

Reverse transcriptases were incubated in 1X reaction buffer. At the indicated time points (5 to 240 minutes), enzyme activity was determined in a standard activity assay.