SMARTScribe Reverse Transcriptase

SMARTScribe RT exhibits exceptional sensitivity

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 105–110 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.

SMARTScribe RT is more sensitive than the competition

SMARTScribe RT is more sensitive than the competition. Human Universal To­tal RNA was serially diluted 10X (from 1 μg 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 manufacturer’s 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 μg. 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.

Comparison of SMARTScribe RT and SMART MMLV RT

Comparison of SMARTScribe RT and SMART MMLV RT.