Abstract: The present invention provides a stable and efficient method for stimulating RNAi-related gene silencing effects. The present invention also provides a fast, simple and specific method for generating amplified cDNA-aRNA hybrids whose quantity and quality are high enough to be used in specific gene silencing transfection. This improved RNA-polymerase cycling reaction (RNA-PCR) relies upon a thermocycling procedure of in-vitro transcription and reverse transcription to bring up the amount of DNA-RNA hybrids up to two thousand folds within one round of the reaction. The resulting cDNA-aRNA product is useful for silencing either endogenous or exogenous gene expression in transfected cells.

Abstract: The present invention provides a fast, simple and reliable polymerase thermocycling reaction procedure for the linear amplification of nucleic acid sequences from cellular RNAs or genomes or both. The principle of this polymerase thermocycling reaction method relies upon the thermal cycling steps of promoter-linked nucleic acid template synthesis and in-vitro transcriptional amplification to bring up the amount of desired nucleic acid sequences up to two thousand fold within one cycle of the above procedure. Neither RNase H activity nor alkaline degradation is used in order to protect the RNA sequences of resulting amplified products which are ready for further applications, such as genechip/microarray analysis, in-vitro translation reaction and induction of RNA interference. Without the preferential amplification drawback of PCR/RT-PCR methods, the accuracy and resolution of current molecular biology analysis and diagnosis can be significantly improved by the present invention.

Abstract: The present invention provides a simple, specific and nontoxic gene knock-out method by formation of covalently bonding between modified probes and targeted sequences. When covalently modified first strand probes are hybridized with a second strand of targeted gene transcripts, certain modified bases of said first strand will interact with natural bases of said second strand to form covalent bonds by which the translation of said second strand is inhibited. Because the hybridization of said two strands generates covalent base-pairing only between their complementary homology region(s), such specificity increases the targeting efficiency of a gene knock-out system.