63 results about "Amplicon sequencing" patented technology

A method for identifying an RNA form of a bacteria, comprising reverse transcribing RNA material; conducting PCR using primers for a first highly conserved genetic sequence generic of the bacteria; conducting nested PCR using primers for a second highly conserved genetic sequence within the first genetic sequence of the bacteria; and identifying the bacteria based on unconserved amplified sequences linked to the conserved sequences.

The invention discloses a construction method of an amplicon library for detecting low-frequency mutation of a target gene. The construction method of the amplicon library for detecting the low-frequency mutation of the target gene, provided by the invention, comprises the following steps: 1) designing and synthesizing a Barcode primer F1, an upstream primer F2, a downstream outer primer R1 and adownstream inner primer R2; 2) carrying out further PCR (Polymerase Chain Reaction) amplification on a cfDNA (cell-free Deoxyribonucleic Acid) of a sample to be detected by utilizing the Barcode primer F1, the upstream primer F2, the downstream outer primer R1 and the downstream inner primer R2, so as to obtain an amplified product, namely a DNA library for amplicon sequencing. By adopting the method disclosed by the invention, a tissue sample can be detected and different regions of free DNAs in samples including blood, urine, cerebrospinal fluid and the like can be rapidly, conveniently, sensitively and specifically subjected to target amplification and mutation which is as low as a 0.1 percent level is efficiently detected; the experiment operation is greatly simplified, the library loss and pollution are effectively avoided, the cost is remarkably reduced and the efficiency is improved.

A method to correct amplification deviation in amplicon sequencing is disclosed. Through steps of amplifying the target nucleic acid to obtain the amplicon coverage of the target nucleic acid, calculating the amplicon coverage ratio between the target nucleic acid in each test genomic region and the target nucleic acid in the reference genomic region, removing outliers, applying a formula to normalize the amplicon coverage ratio, calculating the differences between parameters of the test genomic region amplicon and the reference genomic region amplicon, and applying another formula to fit thedata, and other steps, a regression parameter obtained by fitting is used for correcting the amplification deviation to obtain a normalized amplicon coverage ratio after removing the amplification deviation, thereby eliminating the amplification deviation caused by experimental factors during the multiple PCR amplification process. Elimination of the amplification deviation facilitates accurate calculation of the copy number for a target genomic region, thus enabling detection of minor copy number variation using amplicon sequence data.