RT-PCR 对照引物
品牌	Code No.	包装量	价格
Clontech	639014	25 Rxns	2,134 元

SMARTScribe Reverse Transcriptase

SMARTScribe Reverse Transcriptase is a high-performance enzyme that allows unbiased cDNA synthesis, mRNA amplification, and library construction from any RNA transcript. SMARTScribe RT is a modified Moloney Murine Leukemia Virus Reverse Transcriptase that generates long, full-length cDNA (up to 14.7 kb) and amplifies rare transcripts, while preserving the relative transcript proportions of the original RNA sample. It has been specially formulated for use with all of our SMART Kits.

Our proprietary purification process and rigorous quality control standards ensure that virtually all contaminating nucleases have been removed from SMARTScribe RT.

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

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 105–10 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.