39results about How to "Increased sequencing depth" patented technology

The present invention discloses probes and uses thereof. The group of the probes specifically recognize at least one part of a FLG gene coding region, and meet at least one condition selected from the following conditions that (1) the length of the probe is 75 bp; (2) the probe specifically recognizes the sequence between the upstream 10 bp and the downstream 10 bp of the FLG gene coding region; (3) the multiplier of the probe specifically recognizing the region having the GC content of higher than 0.6 and lower than 0.3 is more than 2; (4) the melting temperatures of the probe and the target sequence is 60-10 DEG C, preferably 80 DEG C; (5) the probe does not contain a hairpin structure; (6) the probe is matched with two sites on a reference genome at the most; and (7) the window sliding size is 10 bp during the probe selection. According to the present invention, the group of the probes provides good capture specificity, high sensitivity and high coverage for at least one part of the specifically-recognized FLG gene coding region, and can be effectively used for the capture detection of the FLG gene coding region.

A mitochondrial genomic library based on high-throughput sequencing and a method for constructing the library. The method comprises the following steps: S1. performing a one-step PCR amplification of total mitochondrial DNA from total DNA; S2. fragmenting the total mitochondrial DNA to obtain a DNA fragment; S3. performing end repair on the DNA fragment to obtain a blunt-end DNA fragment; S4. A-tailing a 3' end of the blunt-end DNA fragment to obtain an A-tailed DNA fragment; S5. adding a linker on the 3' end of the A-tailed DNA fragment to obtain a DNA fragment with linkers; S6. performing LM-PCR on the DNA fragment with linkers; and S7. performing PCR on an LM-PCR product. The method for constructing the mitochondrial genomic library performs PCR amplification on total DNA to obtain total mitochondrial DNA, thereby ensuring accurate amplification of a human mitochondrial genome and improving the accuracy and precision of mutation detection by preventing contamination or mixing of a homologous sequence from a human genome.

The present invention relates to the field of biomedical technology, in particular to a method for detecting tumor mutation load and a prediction model; it includes the following steps: S1: extracting DNA from tumor tissue and blood samples to be tested; S2: constructing target regions for tumor-related genes Targeted capture sequencing library; S3: Using a high-throughput sequencing platform to obtain the original off-machine data; S4: Obtain the tumor mutation load of the sample through the detection process. The purpose of the present invention is to provide a method for detecting tumor mutation load and a prediction model, through which the detection and calculation of gene mutations can achieve the same results as the TMB detection of whole exome sequencing, so as to improve Accuracy of Tumor Mutational Burden Estimation.

The invention relates to the field of molecular biology detection, in particular to a primer, a kit, a model and a construction method for the methylation of cervical cancer-related genes. The methylation primer is designed, and cervical cell DNA is extracted, and the cervical cell DNA is methylated. Methylation treatment, using a PCR amplification kit to perform the first PCR amplification and purification on the cervical cell DNA after methylation treatment, and performing the second PCR amplification and purification on the treated product to obtain a library. The library was subjected to high-pass sequencing on the next-generation sequencing platform for bioinformatics analysis. Based on cervical cancer samples and normal samples, a methylation score was calculated for each PCR amplification region as an independent variable, and the correlation between cervical cancer and cervical cancer was constructed by logistic regression. Gene methylation evaluation model; the present invention builds a database through multiple PCR methylation, and obtains the methylation status of the sample through high-throughput sequencing. The sequencing depth is high, the time is short, and the cost is low. According to the assessed risk, it is decided whether to treat or observe , can effectively reduce overtreatment.