SMARTScribeTM Reverse Transcriptase

Description

SMARTScribe Reverse Transcriptase is a modified Moloney Murine Leukemia Virus Reverse Transcriptase that synthesizes full-length cDNA from rare or long transcripts. SMARTScribe RT preparations are exceptionally pure, with all contaminating nucleases removed. The enzyme is useful for a wide range of applications, and has been specially formulated for use with all of our SMART Kits. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back Back SMARTScribe RT exhibits exceptional sensitivity.RT-PCR assays were performed using SMARTScribe RT and synthetic RNA. The synthetic RNA was serially diluted 10X to obtain 105110 copies per sample; diluted template was then spiked into 50 ng of total RNA from HeLa cells to increase complexity. First-strand cDNA was synthesized with SMARTScribe RT, then amplified with Advantage 2 DNA Polymerase Mix in 36 cycles of PCR using primers specific for a 350-bp amplicon. The PCR reactions were then analyzed by agarose gel electrophoresis. SMARTScribe was able to generate single-stranded cDNA from as few as 10 copies of synthetic RNA. Lane M: 100 bp DNA size marker. Back SMARTScribe RT is more sensitive than the competition.Human Universal Total RNA was serially diluted 10X from 1 mug to 0.1 pg , then reverse transcribed using either SMARTScribe RT Panel A , Competitor P Panel B or Competitor Q Panel C . Each RT reaction was performed according to the enzyme manufacturers specifications. One microliter of each RT reaction was then used in PCR reactions with primers specific for a 300-bp portion of the 5 end of the beta-actin gene. The samples were analyzed by agarose gel electrophoresis. The amount of template RNA used in each reaction is as follows: Lane 1: 1 mug. Lane 2: 100 ng. Lane 3: 10 ng. Lane 4: 1 ng. Lane 5: 100 pg. Lane 6: 10 pg. Lane 7: 1 pg. Lane 8: 0.1 pg. SMARTScribe RT exhibited superior sensitivity at all of the template concentrations tested. Back