Synthetic siRNA generally has a two deoxythymidine nucleotide (dTT) overhang at its 3’ ends. The Synthetic siRNA Quantitation Core Kit adds polydeoxyadenine (poly dA) residues to the 3’ dTT overhangs and then carries out a reverse transcription reaction with a specially-designed oligo dT primer, allowing preparation of complementary DNA from the synthetic siRNA. High-sensitivity siRNA quantitation of synthetic siRNA can be achieved by using the prepared cDNA as the template in SYBR Green I real time PCR with SYBR Premix Ex Taq II (Tli RNase H plus) or a similar product.
Takara's Synthetic siRNA Quantitation Core Kit is designed for siRNA quantitation when used in combination with real time PCR analysis to assay residual siRNA in total RNA samples extracted from cells, tissues, blood and other specimens. There are two kits available, the Synthetic siRNA Quantitation Core Kit (Cat. #6142) and the Synthetic siRNA Quantitation Kit (with SYBR Premix Ex Taq II Tli) (Cat. #RR861A containing Cat. #6142 and Cat. #RR820A).
- This kit is designed for siRNA quantitation of molecules containing 3' d(TT) overhangs.
- For synthetic siRNA with 3ʼoverhangs other than d(TT), prepare a specific oligo dT primer containing the modified sequence.
- This product does not work with synthetic siRNAs that carry no 3’ overhangs or with RNA 3’ overhangs.