54 results about "Gene splicing" patented technology

The invention discloses a one-step seamless, non-homologous and multi-fragment gene splicing method. The steps of DNA fragment phosphorylation, DNA fragment bridging and exponential type amplification of a ligation product are finished in one pipe through thermal cycling similar to PCR. The method has the following advantages: (1) ligation reaction is finished by one step, so that high cost caused by using a phosphorylation primer is avoided; (2) the concentration of DNA, the concentration of a bridging primer and components and a proportion of a buffer solution are optimized, and the ligation reaction with high reliability and good repeatability is reached; (3) DNA fragments are seamlessly ligated with one another without introduction of excess nucleotide; (4) non-DNA sequence dependency is realized; (5) the multi-fragment gene splicing ability is efficient; (6) the ligation product is amplified while ligation, so that the concentration of a product is greatly increased, and efficient conversion and subsequent experiment needs are guaranteed; and (7) the cost is low compared with that of the conventional restriction enzyme ligation method.

Disclosed are systems and methods for simultaneous detection of DNA and RNA genetic alterations comprising gene splicing variants, mutations, indel, copy number changes, fusion and combination thereof, in a biofluid sample from the patient without physically separating RNA from DNA. The systems and methods are similarly applicable to the simultaneous detection of DNA and RNA genetic alterations in solid tissues comprising gene splicing variants, mutations, indel, copy number changes, fusion and combination thereof. The present method utilized a barcoding method for analysis. The streamlined methods improve the simplicity, quantification accuracy and detection sensitivity and specificity of non-invasive detections of biomarkers.