6results about How to "Improve Sequencing Accuracy" patented technology

The invention relates to the technical field of molecular diagnosis, in particular to an F8 gene mutation detection probe group and a kit based on PacBio liquid phase hybridization capture. The probe group comprises sequences as shown in SEQ ID NO. 1 to SEQ ID NO. 313; the F8 gene specific probe is designed, a PacBio sequencing platform is adopted, the type, position and breakpoint of F8 gene mutation can be more accurately determined, the risk of missed diagnosis and misdiagnosis is reduced, when complex variations such as inversion of intron 22 and inversion of intron 1 of the F8 gene are detected, the position of the inverted breakpoint can be clearly recognized through PacBio sequencing, the detection period is shortened, and the detection efficiency is improved. And a way is provided for clinical diagnosis of hemophilia A.

The present invention relates to a method for discriminating between live and dead microbes in a sample, by discriminating between transcriptionally-active and inert microbial nucleic acid sequences in the sample. In particular, the method according to the present invention is based on the comparison of levels of nucleotide substitution in a sample cultured in presence of an RNA-labelling agent. The present invention further relates to a diagnosis method of microbial infections in a subject; and to methods of assessing the risk of contamination of a sample, implementing the method for discriminating according to the present invention.

This invention discloses a CoolMPS antibody mutant that specifically binds to a blocking-modified cytosine deoxynucleotide and its applications. The invention provides an antibody (referred to as an antibody mutant or modified antibody) that binds to a target, obtained by mutating the variable region of the heavy chain of the antibody to be modified; the mutation is located at: amino acid residue 2 of CDR1; amino acid residue 2 of CDR2; amino acid residue 15 of FR1; amino acid residue 6 of FR3; amino acid residue 21 of FR3; amino acid residue 30 of FR3; amino acid residue 38 of FR3; the target is a blocking-modified cytosine deoxynucleotide. The antibody provided by this invention can be used in basic scientific research, biosensor technology, gene sequencing, and other fields, and has broad application prospects.

The invention relates to a single-chain connector, a method for detecting the tail end of a nucleic acid fragment, a kit and application of the single-chain connector. The single-chain connector disclosed by the invention can be simultaneously connected with the same-side tail end of a to-be-detected double-chain nucleic acid fragment, a protruding tail end sequence of double-chain nucleic acid can be efficiently and accurately captured, and the motif and position information of the tail end of the double-chain nucleic acid can be favorably and accurately identified through sequencing analysis.

The invention discloses a single-ended multiplex PCR (Polymerase Chain Reaction) library building method for pathogenic microorganisms, relates to the technical field of microbial gene detection, and in particular relates to the single-ended multiplex PCR library building method for the pathogenic microorganisms. The method comprises the following steps: designing primers, downloading a reference genome from NCBI (National Center of Biotechnology Information), sliding to generate a kmer sequence, comparing to an nt reference library by using blast, filtering non-specific kmer, screening proper primer sequences by using MFEprimer, and selecting a pair of inner and outer side primer sequences from each species; pCR amplification is carried out on the basis of the outer side primer by using the inner side primer, the specificity is enhanced, and then the primer is synthesized according to a self-designed primer sequence. After the technical scheme is adopted, the method has the beneficial effects that the pathogenic microorganisms are detected by using an ultra-high-weight PCR technology and a second-generation high-throughput sequencing technology, and a library can be established by adding 10ng of a sample; multiple single-ended primers can be directly mixed for use, and the concentration of the single primers does not need to be adjusted; the RNA sample and the DNA sample can be used for building a database together, so that the experimental operation is simplified.

The invention discloses a primer set and a detection method for detecting different types of a human rhinovirus, and relates to the technical field of microbial gene detection, the rhinovirus is a single-chain RNA virus, and a 0bp-650bp non-coding region needs to be selected in 7000bp. The primer set and the detection method have the advantages that the primer set is designed for the human rhinovirus, various types of the human rhinovirus can be effectively detected, and the detection accuracy is high. According to the present invention, the pathogenic microorganisms are detected by using the ultra-high heavy PCR technology and the second-generation high-throughput sequencing technology, the library can be established by adding 10 ng of the sample, all the primers can be directly mixed for use, the concentration of the single primer does not need to be adjusted, the experiment operation is simple and convenient, the whole process operation can be completed within 5 h, the orthogonal combination between the Index can be performed by using the double-end Index, the Index diversity is enriched, and the detection efficiency is high; according to the method, the final library mixing quantity of each lane is increased, the Index hopping effect is effectively reduced, the data is accurately split, and the sequencing accuracy is improved.