Ultrapure SMART MMLV Reverse Transcriptase for RT-PCR

SMART MMLV Reverse Transcriptase is an ultra-pure Moloney Murine Leukemia Virus Reverse Transcriptase designed to virtually eliminate contaminating RNases and other nucleases that are often introduced into the RT reaction with the enzyme. SMART MMLV RT helps prevent degradation of template RNA, while allowing the synthesis of high quality, full-length cDNA (up to 11.7 kb) from almost any transcript.

SMART MMLV RT synthesizes long, full-length cDNAs. SMART MMLV RT was used to reverse transcribe first-strand cDNA from Human Brain Total RNA using oligo(dT)18 primers (Panel A). The resulting single-stranded cDNA was then serially diluted fivefold (corresponding to 25 ng–1.6 pg of RNA) and analyzed by qPCR using QTaq DNA Polymerase Mix and gene-specific primers that amplified a 226 bp region near the 5' end of NF1 (Panel B). The successful amplification of this region indicates that SMART MMLV RT was able to reverse transcribe at least 11.7 kb of the NF1 gene.

Comparison of SMARTScribe RT and SMART MMLV RT

SMART MMLV RT is ideal for SMART mRNA amplification. Full-length sense mRNA was amplified from 0.1 μg and 1 μg of Human Placenta Total RNA using our SMART mRNA Amplification Kit (which uses T7 RNA Polymerase). The amplified mRNA was then analyzed on a denaturing agarose gel. Lane 1: RNA ladder; Lanes 2 & 3: mRNA amplified from 1 μg of total RNA; Lanes 4 & 5: mRNA amplified from 0.1 μg of total RNA; Lane 6: Negative Control (–RNA).

SMART MMLV RT has less RNase activity than the competition. An RNA ladder was incubated with 200 units of either SMART MMLV RT (Lanes 1 & 2), or another supplier’s MMLV RT (Lanes 3 & 4) and AMV RT (Lanes 5 & 6) at 37°C for 1 hr, and then analyzed by formaldehyde/agarose gel electrophoresis. Lane 7: negative control (buffer only, no enzyme). Lane 8: positive control.