Iscript cdna synthesis kit protocol, related categories
The simplicity of the protocol allows me to run multiple reactions for first-strand cDNA synthesis in PCR tubes or multi-well plates. I found it to be well-suited for standard PCR with post-amplification analysis in agarose gels with ethidium bromide staining, as well.
It is designed to enable two-step RT-PCR analysis with high sensitivity over a wide range of RNA sources and optimized for amplification of targets less than 1 kb in size.
Technique de dissertation en droit inhibitor, a blend of oligo dT and random hexamer primers and dNTPs are all included in the 5x transcription mix, which minimizes both pipetting and time requirements for setup. The documentation is succinct, but sufficient.
The kit allows researchers to run extremely convenient and consistent synthesis kit protocol transcription reactions, as everything is assembled in the tubes at once without requiring separate steps for heat denaturation, chilling or primer annealing minimizing the chance of cross-contamination.
The kit has shown great sensitivity for low abundance and complex targets, even when the initial RNA amount is significantly less than 1 ug.
The competitively priced kit is provided in two sizes 25 or reactions. Following RNA addition, the plates are sealed and cDNA is synthesized in a thermal cycler without any additional attention requirement. Once the reactions are mixed and spun, the tubes are incubated for 5 min at 25C, for 30 min at 42C and for 5 min at 85C.
I used this kit to make cDNA from total RNA isolated from established cell lines human lung epithelial cell line A and mouse lymphoma cell line EL4mouse primary T lymphocytes and tissues lungs and livers.
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