Thermo Scientific™ Maxima H Minus First Strand cDNA Synthesis Kit

Description

• Thermo Scientific Maxima H Minus First Strand cDNA Synthesis Kit, available with or without dsDNase, is a complete system for highly efficient synthesis of first strand cDNA
• The kit uses the Maxima H Minus Reverse Transcriptase (RT), which is an advanced enzyme derived by in vitro evolution of MMLV RT
• The enzyme features the highest thermostability among the derivatives of MMLV RT and lacks RNase H activity
• 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 • First Strand cDNA synthesis for RT-PCR • Construction of cDNA libraries • Generation of probes for hybridization • Antisense RNA synthesis Contents • Maxima H Minus Enzyme Mix • Oligo(dT) 18 and Random hexamer primers • 5X RT Buffer • dNTP Mix • Nuclease-free water First Strand Synthesis Kit with dsDNase also contains • dsDNase and 10X dsDNase Buffer 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.