A method and a kit for detecting genome editing and application thereof belongs to the field of genome editing efficiency detection, and the getPCR method for determining genome editing efficiency includes quantifying wild-type DNA in a genome to be tested and calculating the percentage of the wild-type DNA to determine the genome editing efficiency. The method has been proved to have good detection accuracy and simple operation, and can be applied to all genome editing methods to quantify genome editing efficiency and screen single-cell clones.

Techniques are provided for generating an array-specific range of Tm values to be used for calling a sample in a given array positive or negative for a target nucleic acid sequence. A sample well in an array is provided with a control sample containing a control nucleic acid sequence. The control sample is amplified by thermal cycling the sample well. A Tm value for the control sample is identified and compared to an expected Tm value for the control nucleic acid sequence to calculate a relationship between the identified control Tm value and the expected control Tm value. By applying this relationship to an expected Tm value for a target nucleic acid sequence, an array-specific range of Tm values for the target nucleic acid sequence is generated and can be used for calling an experimental sample in the same array positive or negative for the target nucleic acid sequence.

Techniques are provided for generating an array-specific range of Tm values to be used for calling a sample in a given array positive or negative for a target nucleic acid sequence. A sample well in an array is provided with a control sample containing a control nucleic acid sequence. The control sample is amplified by thermal cycling the sample well. A Tm value for the control sample is identified and compared to an expected Tm value for the control nucleic acid sequence to calculate a relationship between the identified control Tm value and the expected control Tm value. By applying this relationship to an expected Tm value for a target nucleic acid sequence, an array-specific range of Tm values for the target nucleic acid sequence is generated and can be used for calling an experimental sample in the same array positive or negative for the target nucleic acid sequence.

Disclosed are methods for determining at least one sequence of interest of a fetus of a pregnant mother. In various embodiments, the method can determine one or more sequences of interest in a test sample that comprises a mixture of fetal cellular DNA and mother-and-fetus cfDNA. In some embodiments, methods are provided for determining whether the fetus has a genetic disease. In some embodiments, methods are provided for determining whether the fetus is homozygous in a disease causing allele when the mother is heterozygous of the same allele. In some embodiments, methods are provided for determining whether the fetus has a copy number variation (CNV) or a non-CNV genetic sequence anomaly.

Disclosed are methods for determining at least one sequence of interest of a fetus of a pregnant mother. In various embodiments, the method can determine one or more sequences of interest in a test sample that comprises a mixture of fetal cellular DNA and mother-and-fetus cfDNA. In some embodiments, methods are provided for determining whether the fetus has a genetic disease. In some embodiments, methods are provided for determining whether the fetus is homozygous in a disease causing allele when the mother is heterozygous of the same allele. In some embodiments, methods are provided for determining whether the fetus has a copy number variation (CNV) or a non-CNV genetic sequence anomaly.

In one embodiment, provided are a method for analyzing at least one nucleic acid that can conveniently and highly accurately analyze even a very small number of analyte at least one nucleic acid. In one embodiment, the present invention relates to a method for analyzing at least one nucleic acid, comprising: a library preparation step of preparing a library comprising at least one standard nucleic acid of specific copy number(s) and at least one analyte nucleic acid in a same system; a calibration curve data generation step of generating calibration curve data based on the copy number(s) of the at least one standard nucleic acid of specific copy number(s); and an analyte nucleic acid analysis step of identifying at least one nucleotide sequence of the analyte nucleic acid while identifying the number(s) of the at least one nucleotide sequence of the at least one analyte nucleic acid using the calibration curve data.

In one embodiment, provided are a method for analyzing at least one nucleic acid that can conveniently and highly accurately analyze even a very small number of analyte at least one nucleic acid. In one embodiment, the present invention relates to a method for analyzing at least one nucleic acid, comprising: a library preparation step of preparing a library comprising at least one standard nucleic acid of specific copy number(s) and at least one analyte nucleic acid in a same system; a calibration curve data generation step of generating calibration curve data based on the copy number(s) of the at least one standard nucleic acid of specific copy number(s); and an analyte nucleic acid analysis step of identifying at least one nucleotide sequence of the analyte nucleic acid while identifying the number(s) of the at least one nucleotide sequence of the at least one analyte nucleic acid using the calibration curve data.

The present invention provides an improved quantitative PCR method (qPCR method) for measuring a quantity of a target gene included in a human cell, the method enabling a more accurate analysis of cell kinetics in cell therapy, for example. The qPCR method of the present invention includes a step of adding, to a biological sample including the human cell, a predetermined quantity of exogenous DNA that does not cross over with the target gene as an external standard for correcting a measured quantity of the target gene.

The present invention provides an improved quantitative PCR method (qPCR method) for measuring a quantity of a target gene included in a human cell, the method enabling a more accurate analysis of cell kinetics in cell therapy, for example. The qPCR method of the present invention includes a step of adding, to a biological sample including the human cell, a predetermined quantity of exogenous DNA that does not cross over with the target gene as an external standard for correcting a measured quantity of the target gene.

Provided herein is a nucleotide cassette comprising an inducible promoter, a nucleotide sequence that corresponds to at least one single stranded RNA diagnostic target, a nucleotide sequence that encodes artemin, a molecular switch and a nucleotide sequence that encodes a DNAse enzyme and is under control of the molecular switch, wherein the single stranded RNA diagnostic target is a sequence detected by a molecular diagnostic assay. In some embodiments the nucleotide cassette can be used to obtain an RNA expression product. Also provided are vectors and cells comprising the nucleotide cassette or the RNA expression product thereof. The nucleotide cassette can further be used to obtain a diagnostic control composition comprising a non-pathogenic recombinant bacterium having a modified genetic content comprising the nucleotide cassette and to methods of producing such recombinant bacteria.