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Maxima Reverse Transcriptase

Kód produktu: EP0743 Kód výrobce: EP0743 Kód dodavatele: {E1385276-E6CB-469E-8732-15DE57F7CBA2} Výrobce: Life Technologies Czech Republic s.r.o.
24 103,20 Kč
19 920,00 Kč bez DPH
do týdne

An advanced enzyme developed through in vitro evolution of M-MuLV RT.

Zobraz detailní popis

Detailní popis

High yields of cDNA over a broad temperature range

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 (see supporting data). 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

Includes

  • Maxima Reverse Transcriptase (200 U/µL)
  • 5X RT Buffer

Note

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

5x RT Buffer 250 mM Tris-HCl (pH 8.3 at 25°C), 375 mM KCl, 15 mM MgCl2, 50 mM DTT
Definition of Activity Unit
  • One unit of the enzyme incorporates 1 nmol of dTMP into a polynucleotide fraction (adsorbed on DE-81) in 10 minutes at 37°C.
  • Enzyme activity is assayed in the following mixture: 50 mM Tris-HCl (pH 8.3), 4 mM MgCl2, 10 mM DTT, 50 mM KCl, 0.5 mM dTTP, 0.4 MBq/mL [3H]-dTTP, 0.4 mM polyA oligo(dT)12-18.
Hazardous No
Inactivation Inactivated by heating at 85°C for 5 minutes
Inhibition Metal chelators, inorganic phosphate, pyrophosphate, and polyamines
Quality Control
  • The absence of endodeoxyribonucleases, exodeoxyribonucleases, phosphatases, and ribonucleases confirmed by appropriate quality tests.
  • Functionally tested in first strand cDNA synthesis.
Source E.coli cells carrying an engineered pol gene fragment of Moloney Murine Leukemia Virus.
Storage Buffer 50 mM Tris-HCl (pH 7.5), 0.1 M NaCl, 1 mM EDTA, 5 mM DTT, 0.1% (v/v) Triton X-100, and 50% (v/v) glycerol
Storage Condition -20 C

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