PrimeScript Reverse Transcriptase is a modified MMLV (Moloney Murine Leukemia Virus) RTase. This enzyme has extremely high extension capability and can synthesize long first-strand cDNA efficiently. Even for difficult templates, including RNAs with complex secondary structure, it is possible to synthesize first-strand cDNA at the normal reverse transcription temperature (42℃) with this enzyme. It is not necessary to perform the RT reaction at higher temperatures (a condition that may cause RNA degradation). This enzyme is suitable for preparation of long cDNAs, for construction of cDNA libraries that include full-length cDNA, etc.
Features
Strong strand-displacement and extension capability: capable of synthesizing long and full-length cDNA molecules (up to 12 kb)
Preparation of cDNA at 42°C results in lower background and higher cDNA yields because the lower temperature does not promote RNA degradation
Outstanding accuracy: lowest error rate among five commercially available reverse transcriptases tested
Highly specific: low rates of non-specific annealing, even on incompletely denatured RNA
Works on challenging templates: excellent results even with GC-rich templates and templates with high levels of secondary structure
Applications
RT-PCR First-strand cDNA synthesis cDNA probe preparation Synthesis of cDNA libraries with a high proportion of full-length cDNAs
Unit definition
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
