This Reverse Transcriptase has exceptionally strong strand displacement activity and enables efficient preparation of cDNA up to 12 kb in length. It is robust, versatile and well-suited for applications requiring full-length cDNA such as preparation of cDNA libraries and other techniques involving first strand cDNA synthesis (RT-PCR, preparation of cDNA probes, real-time quantitative RT-PCR). It can be used for performing a reverse transcription reaction with any RNA template including GC-rich templates and RNAs with high levels of secondary structure. This enzyme is a modified, recombinant MMLV (Moloney Murine Leukemia Virus) reverse transcriptase and is verified to be RNase H Minus. Because of the excellent extension capability of PrimeScript Reverse Transcriptase, preparation of cDNA can be performed at a lower temperature (42°C), decreasing the risk of RNA degradation that can occur during conventional reactions performed at higher temperatures.
RT, Recombinant (Moloney Murine Leukemia Virus)
Moloney Murine Leukemia Virus
RT-PCR; First strand cDNA synthesis; cDNA probe preparation; Synthesis of cDNA libraries with a high proportion of full-length cDNAs.
1. Strong strand displacement and extension capability: Capable of synthesizing long and full-length cDNA molecules (up to 12 kb); 2. Preparation of cDNA at 42°C results in lower background and higher cDNA yields because the lower temperature does not promote RNA degradation; 3. Outstanding accuracy: Lowest error rate among five commercially available reverse transcriptases tested; 4. Highly specific: Low rates of non-specific annealing, even on incompletely denatured RNA; 5. Works on challenging templates: Excellent results even with GC-rich templates and templates with high levels of secondary structure 6. Highly sensitive: use less of your precious RNA samples; 7. Demonstrated success with reverse transcription reaction times of 30 min. (60 min. for longer transcripts).
One unit is the amount of the enzyme that incorporates 1 nmol of [3H]dTTP in 10 minutes at 37°C, with poly (rA), oligo (dT) 12-18 as the primer-template.
5X Buffer (for cDNA synthesis) 250 mM Tris-HCl, pH 8.3 375 mM KCl 15 mM MgCl2 Reaction mixture for unit definition: 50 mM: Tris-HCl, pH 8.3 75 mM: KCl 8mM: MgCl2 10 mM: DTT 20 ug/mL: (ra)n (dT)12-18 0.5 mM: [3H]dTTP 0.1%: NP-40 Storage Buffer Composition 200 mM Tris-HCl, pH 7.8 100 mM NaCl 1 mM EDTA 1 mM DTT 50% Glycerol (v/v)
Reverse transcriptase; RT