Patent classifications
C12Q2600/166
Control compositions and methods for sequencing
The invention relates to control compositions for sequencing and for chemical analyses, such as analytical chemistry analyses. More particularly, the invention relates to control compositions for sequencing and for chemical analyses having at least one barcode sequence fragment and at least one universal sequence fragment, and to methods of their use.
QUALITY CONTROL TEMPLATES ENSURING VALIDITY OF SEQUENCING-BASED ASSAYS
Embodiments of a method and/or system can include generating a set of quality control template (QCT) molecules; determining a set of QCT sequence read clusters based on the set of QCT molecules, such as based on variation regions of the set of QCT molecules; and based on the set of QCT sequence read clusters, determining a sequencing-related parameter, such as a contamination parameter and/or molecule count parameter, associated with the at least one of sequencing library preparation and sequencing.
COMPOSITION AND REAGENT KIT FOR EARLY DETECTION OF CERVICAL HIGH-GRADE LESIONS AND CERVICAL CANCER
The invention discloses a composition and kit for early detection of high-grade cervical lesions and cervical cancer, wherein the composition for early detection of high-grade cervical lesions and cervical cancer includes methylation primers, a probe corresponding to methylated sites and methylation blocking primers for FAM19A4 gene; methylation primers, a probe corresponding to methylated sites and methylation blocking primers for JAM3 gene; methylation primers, a probe corresponding to methylated sites and methylation blocking primers for PAX1 gene; and 1 pair of primers and a probe corresponding to methylated sites for internal reference gene GAPDH. The methylated sites in FAM19A4, JAM3 and PAX1 genes are accurately detected using multiple multi-channel fluorescence and blocking techniques through accurate recognition between specific primers and probes and methylated sequences, full release of methylated templates under the action of multiple blocking primers and optimized special methylation DNA polymerase.
PCR controls
The present invention provides a method of quantification of a target nucleic acid, using at least any two of the genes SYT10, EPHA3, PLEKHF1 and KBTBD4 as control genes. In particular, the combination of the genes SYT10, EPHA3, PLEKHF1 and KBTBD4, known as the 4Plex, is provided as a control for nucleic acid quantification. The 4Plex has particular utility as a control for nucleic acid quantification by methylation-specific droplet digital PCR.
SAMPLE FOR EVALUATING PERFORMANCE OF GENETIC TESTING APPARATUS, METHOD FOR PREPARING SAID SAMPLE, AND DEVICE FOR EVALUATING PERFORMANCE, METHOD FOR EVALUATING PERFORMANCE, PROGRAM FOR EVALUATING PERFORMANCE, AND APPARATUS FOR EVALUATING PERFORMANCE OF GENETIC TESTING APPARATUS
A sample for evaluating performance of a genetic testing apparatus includes, a nucleic acid A; and a nucleic acid B, in which the nucleic acid A and the nucleic acid B have mutually different sequences. The nucleic acid A contains a specific number of molecules, and the nucleic acid B contains a number of molecules higher than the number of molecules of the nucleic acid A. A ratio A/B of the number of molecules of the nucleic acid A with respect to the number of molecules of the nucleic acid B is specified.
Compositions and methods for detecting a biological contaminant
Provided are compositions and methods useful to the determination of whether a microbial contaminant is present in a biological therapeutic production process. Specifically, an artificial positive amplification control plasmid and unique quantitative PCR detection probe are provided, which enables the rapid and real-time detection of a false positive result.
Synthetic nucleic acid spike-ins
This disclosure provides methods for determining relative abundance of one or more non-host species in a sample from a host. Also provided are methods involving addition of known concentrations of synthetic nucleic acids to a sample and performing sequencing assays to identify non-host species such as pathogens. Also provided are methods of tracking samples, tracking reagents, and tracking diversity loss in sequencing assays.
CELL-FREE DNA METHYLATION AND NUCLEASE-MEDIATED FRAGMENTATION
Nuclease activity can affect the methylation level and fragmentation of cfDNA. Certain levels of nuclease activity may be correlated with certain levels of methylation in certain regions. Methylation level in certain genomic regions can be analyzed to classify nuclease activity. Methylation statuses of different genomic regions compared to methylation statuses of other genomic regions can determine a level of a condition (e.g., a disease such as cancer or disorder) in a subject. Nuclease activity can be monitored through analysis of methylation statuses of different sites. The efficacy of a treatment can also be determined using methylation levels at certain genomic regions. The number of fragments from genomic regions that are hypomethylated or hypermethylated in a reference genome can be used to provide information (e.g., fractional concentration) on the sample itself. The size distribution of extrachromosomal circular DNA can also be used to analyze a biological sample. Systems are also described.
NUCLEIC ACID LINKED IMMUNE-SANDWICH ASSAY (NULISA)
Disclosed herein are highly sensitive immunoassays that utilize a capture/release mechanism to reduce non-specific binding and achieve detection with attomolar-level sensitivity. Kits that can be used for carrying out these highly sensitive immunoassays are also disclosed herein.
Single-cell RNA sequencing using click-chemistry
The present disclosure relates to a method of sequencing nascent RNA in a cell. In some embodiments, the nascent RNA is conjugated to DNA using copper-catalyzed azide-alkyne cycloaddition (CuAAC). Methods of the present disclosure can be used to generate genomic libraries of a cell and measure gene expression and enhancer and/or super-enhancer activity.