Maxima H Minus First Strand cDNA Synthesis Kit

This kit allows synthesis of long cDNAs up to 20 kb at elevated temperatures (up to 65°C), superseding other systems' abilities to produce full-length cDNA. Due to increased synthesis rates the reaction can be completed in 30 minutes.

For reverse transcription the kit uses Maxima H Minus Reverse Transcriptase (RT), which is an advanced enzyme derived by in vitro evolution of MMLV RT.

The Maxima H Minus First Strand cDNA Synthesis Kit with dsDNase provides a dramatically simplified workflow that combines genomic DNA elimination and cDNA synthesis into one-tube procedure. The kit contains a novel double-strand-specific DNase (dsDNase) engineered to remove contaminating genomic DNA from RNA preps in two minutes without damage to quality or quantity of RNA. Highly specific dsDNase activity towards double-stranded DNA ensures that single-stranded DNA (such as cDNA and primers) is not cleaved, and dsDNase treated RNA can be directly added to reverse transcription.

Features of the Maxima H Minus First Strand cDNA Synthesis Kit include:

• Increased reaction temperatures—the first strand of cDNA can be synthesized within the 42–65°C temperature range
• High yields of full-length first strand cDNA—with RNA templates up to 20 kb
• Flexible priming—oligo(dT)₁₈, random hexamer or gene-specific primers
• Integrated genomic DNA removal step with dsDNase kit

Applications

• Full-length cDNA synthesis for long transcripts and ORFs
• Reverse transcription at elevated temperatures to reduce effects of secondary structure
• cDNA cloning and construct generation
• RNA-seq & single cell RNA-seq library prep
• Isoform and transcript characterization
• Template-switching-based full-length RNA-seq

Additional information about reaction components

• Maxima H Minus Enzyme Mix contains Maxima H Minus Reverse Transcriptase and RiboLock RNase Inhibitor. RiboLock RNase Inhibitor effectively protects RNA templates from degradation by RNases A, B, and C at temperatures up to 55°C.
• Oligo(dT)₁₈ and random hexamer primers are supplied with the kit. Random hexamer primers bind non-specifically and are used to synthesize cDNA from all RNAs in a total RNA population. The oligo(dT)18 primer selectively anneals to the 3'-end of poly(A) RNA, synthesizing cDNA only from poly(A) tailed mRNA. Gene-specific primers may also be used with the kit to prime synthesis from a specified sequence.
• 10 mM dNTP Mix is a premixed aqueous solution of dATP, dTTP, dCTP, and dGTP.
• Nuclease-free water is provided for reaction set-up and dilution of sample DNA. The absence of endo-, exodeoxyribonucleases, ribonucleases, and phosphatases has been confirmed by appropriate quality tests.