Thermo Scientific Maxima™ Reverse Transcriptase products are designed for consistency and efficiency in cDNA synthesis for RT-PCR and RT-qPCR applications.
Maxima H Minus Reverse Transcriptase is engineered with molecular evolution techniques selecting for mutations that confer dramatically improved thermostability, processivity, and activity rates compared to wild-type M-MuLV enzymes. These features support higher cDNA yields with a variety of templates and improved synthesis from templates with complex secondary structures. In RT-qPCR applications, Maxima H Minus Reverse Transcriptase also enables high efficiency synthesis over a wide range of input template amounts providing sensitive and accurate quantification of cDNA.
Maxima H Minus RT has 50X increased processivity compared with wild type MMuLV reverse transcriptase enzymes. This reverse transcriptase is capable of synthesizing full-length cDNA from a wide range of RNA templates (Figure 2).
Maxima H Minus Reverse Transcriptase efficiently synthesizes cDNA from a wide range of template amounts, has higher yields, and better linearity in cDNA synthesis outperforming other commercially available reverse transcriptases, making it an exceptional choice for RT-qPCR experiments (Figure 3). The premixed solutions in the Thermo Scientific Maxima First Strand cDNA Synthesis Kits help further improve reproducibility and reaction setup time.
Figure 3. Consistently efficient reverse transcription over a wide range of input RNA amounts. Maxima H Minus First Strand cDNA Synthesis kit demonstrates consistently better reverse transcription efficiency than competitor kits. Amplification plots show variation of log (ΔRn) with PCR cycle number. RT-qPCR of human β-2 macroglobulin gene was performed from 10-fold serial dilutions of HeLa total RNA (1 μg to 1 pg). First strand cDNA was generated using the Maxima H Minus First Strand cDNA Synthesis kit and 7 other commercial First Strand cDNA Synthesis kits. cDNA was amplified using TaqMan Universal Master Mix II, with UNG on the Applied Biosystems ViiA7 Real-Time PCR System.
The Maxima H Minus cDNA Synthesis Master Mix maintains high transcription efficiency and good linearity across a broad range of template concentrations (Figure 4). The observed linearity strongly suggests reliable representation of relative quantities of different transcripts when using large or small amounts of input RNA.
The Maxima H Minus cDNA Synthesis Master Mix offers higher efficiency than reverse transcriptases from other suppliers at low and high input RNA amounts (Figure 5). The higher transcription efficiency allows the use of less RNA and accurate detection of less expressed transcripts.
The Maxima H Minus cDNA Synthesis Master Mix shows consistently better efficiency than reverse transcriptase master mixes from other suppliers over a range of 96 target genes in RT-qPCR when normalized to data generated with the Maxima First Strand cDNA Synthesis Kit for RT-qPCR (Figure 6).
The double-strand-specific DNase (dsDNase) enables faster, efficient, and complete gDNA removal compared to conventional DNase I (Figure 7). dsDNase-treated RNA samples show no decrease in RNA integrity or quantity. With the use of dsDNase treatment prior to cDNA synthesis, the risk of sample loss can be minimized.
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