Description

Get highly efficient synthesis of first strand cDNA with the Thermo Scientific™ Maxima™ H Minus First Strand cDNA Synthesis Kit.

The kit uses the Maxima H Minus Reverse Transcriptase (RT), which is an advanced enzyme derived by in vitro evolution of M-MuLV RT. The enzyme features the highest thermostability among the derivatives of M-MuLV RT and lacks RNase H activity. The Maxima H Minus First Strand cDNA Synthesis 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.
The new 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 2 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.

Highlights

• Integrated genomic DNA removal step
• Increased reaction temperatures - the first strand of cDNA can be synthesized within the 42 to 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

Applications

• First Strand cDNA synthesis for RT-PCR
• Construction of cDNA libraries
• Generation of probes for hybridization
• Antisense RNA synthesis

Includes

• Maxima H Minus First Strand cDNA Synthesis Kit contains Maxima H Minus Enzyme Mix, Oligo(dT)₁₈ and Random hexamer primers, 5X RT Buffer, dNTP Mix, and nuclease-free water.
• Maxima H Minus First Strand cDNA Synthesis Kit with dsDNase, in addition to the above listed components, 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. †Note that the name of this product has changed. The previous name is RevertAid Premium First Strand cDNA Synthes

Hazardous: No
Quality Control: Functionally tested in RT-PCR using 100 fg of control GAPDH RNA and GAPDH control primers generate a prominent 496 bp product on 1% agarose gel after ethidium bromide staining.
Storage Condition: -20°C