1006 results about "Nested polymerase chain reaction" patented technology

The present invention relates to the detection of nucleic acid sequence differences using coupled ligase detection reaction and polymerase chain reaction. One aspect of the present invention involves use of a ligase detection reaction coupled to a polymerase chain reaction. Another aspect of the present invention relates to the use of a primary polymerase chain reaction coupled to a secondary polymerase chain reaction coupled to a ligase detection reaction. A third aspect of the present invention involves a primary polymerase chain reaction coupled to a secondary polymerase chain reaction. Such coupling of the ligase detection reaction and the polymerase chain reaction permits multiplex detection of nucleic acid sequence differences.

Clinical management of human cancer is dependent on the accurate monitoring of residual and recurrent tumors. We have developed a method, called personalized analysis of rearranged ends (PARE), which can identify translocations in solid tumors. Analysis of four colorectal and two breast cancers revealed an average of nine rearranged sequences (range 4 to 15) per tumor. Polymerase chain reaction with primers spanning the breakpoints were able to detect mutant DNA molecules present at levels lower than 0.001% and readily identified mutated circulating DNA in patient plasma samples. This approach provides an exquisitely sensitive and broadly applicable approach for the development of personalized biomarkers to enhance the clinical management of cancer patients.

Highly specific and sensitive methods were developed for multiplex amplification of nucleic acids on supports such as microarrays. Based on a specific primer design, methods include five types of amplification that proceed in a reaction chamber simultaneously. These relate to four types of multiplex amplification of a target DNA on a solid support, directed by forward and reverse complex primers immobilized to the support and a fifth type—pseudo-monoplex polymerase chain reaction (PCR) of multiple targets in solution, directed by a single pair of unbound universal primers. The addition of the universal primers in the reaction mixture increases the yield over the traditional “bridge” amplification on a solid support by approximately ten times. Methods that provide multitarget amplification and detection of as little as 0.45-4.5×10−12 g (equivalent to 102-103 genomes) of a bacterial genomic DNA are disclosed.

The invention relates to compositions, methods, and kits for nucleic acid replication, including polymerase chain reaction (PCR) and mutagenesis reactions. A buffer composition is provided which allows higher concentrations of DNA polymerase to be used, resulting in greater yield of amplified product and faster reaction kinetics.

The invention relates to a nucleic acid releasing agent, a nucleic acid PCR (polymerase chain reaction) amplification method and a PCR amplification kit. The nucleic acid releasing agent comprises Tris-HCl, sodium chloride, potassium chloride, Tween 20, Triton X-100, Nonidet P-40 and strong base; the molar concentration of the Tris-HCl is 0.5-500mM, the molar concentration of the sodium chloride is 20-500mM, the volume percentage of the Tween 20 is 0.1-2%, the volume percentage of the Triton X-100 is 0.1-3%, the volume percentage of the Nonidet P-40 is 0.1-3%, the mass concentration of the potassium chloride is 5-8mg / mL, and the mass concentration of the strong base is 2-50mg / mL. The nucleic acid releasing agent has the advantages that RNA-containing samples can directly release RNAs at the room temperature, and can be directly mixed with PCR reaction liquid to achieve PCR amplification, so that amplification can be completed without complex nucleic acid extraction and purification processes.