Provided herein are automated apparatus and methods for the identification of microorganisms in various samples. The disclosure solves existing challenges encountered in identifying and distinguishing various types of microorganisms, including viruses and bacteria in a timely, efficient, and automated manner by library preparation and sequencing.

The present invention discloses markers, primers, probes and a kit for early screening and diagnosis of endometrial cancer. The markers are partial methylated regions in the four genes of CDO1, CELF4, HAND2 and HS3ST2. Detection primers and probes are designed for these methylated regions and to have clasp structures. The kit includes the aforementioned primers and probes. The present invention screens and combines multiple methylated regions to determine the most suitable methylated position for combined diagnosis, which can significantly improve the sensitivity and specificity of the detection for early endometrial cancer. The kit is especially suitable for early screening and diagnosis of endometrial cancer with cervical exfoliated cells as a sample. Even if the DNA template concentration is low, endometrial cancer can be detected. The kit has the advantages of non-invasive sampling, fast detection speed, higher sensitivity and specificity, etc., and can ensure the accuracy of the results to help achieve the purpose of early detection, early diagnosis, and early treatment of endometrial cancer.

The present invention discloses markers, primers, probes and a kit for early screening and diagnosis of endometrial cancer. The markers are partial methylated regions in the four genes of CDO1, CELF4, HAND2 and HS3ST2. Detection primers and probes are designed for these methylated regions and to have clasp structures. The kit includes the aforementioned primers and probes. The present invention screens and combines multiple methylated regions to determine the most suitable methylated position for combined diagnosis, which can significantly improve the sensitivity and specificity of the detection for early endometrial cancer. The kit is especially suitable for early screening and diagnosis of endometrial cancer with cervical exfoliated cells as a sample. Even if the DNA template concentration is low, endometrial cancer can be detected. The kit has the advantages of non-invasive sampling, fast detection speed, higher sensitivity and specificity, etc., and can ensure the accuracy of the results to help achieve the purpose of early detection, early diagnosis, and early treatment of endometrial cancer.

The present invention relates to methods of diagnosing bladder cancer in a patient, involving determining the methylation status of Methylation Variable Positions (MVPs) in DNA from the patient and providing a diagnosis based on methylation status data. The invention also relates to methods of treating bladder cancer comprising providing a diagnosis of bladder cancer by the diagnostic methods defined herein followed by administering one or more anti-cancer agents to a patient. The invention also relates to methylation-discriminatory arrays comprising probes directed to the MVPs defined herein and kits comprising the arrays.

The presently disclosed subject matter provides high-throughput methods for performing genomic DNA methylation assessments. The presently disclosed subject matter further provides methods for diagnosing a subject with a disease and/or disorder, and for determining the prognosis of a subject that has a disease and/or disorder. In certain embodiments, the present disclosure provides a diagnostic method that includes obtaining a biological sample from the subject; determining the methylation status of one or more genomic DNA loci in one or more cells of the biological sample; and diagnosing a disease and/or disorder in the subject, wherein the methylation status of the one or more genomic DNA loci indicates the presence of the disease and/or disorder in the subject.

The presently disclosed subject matter provides high-throughput methods for performing genomic DNA methylation assessments. The presently disclosed subject matter further provides methods for diagnosing a subject with a disease and/or disorder, and for determining the prognosis of a subject that has a disease and/or disorder. In certain embodiments, the present disclosure provides a diagnostic method that includes obtaining a biological sample from the subject; determining the methylation status of one or more genomic DNA loci in one or more cells of the biological sample; and diagnosing a disease and/or disorder in the subject, wherein the methylation status of the one or more genomic DNA loci indicates the presence of the disease and/or disorder in the subject.

The present invention is based on the discovery of sensitive and specific methylation detection by a) controlling excessive DNA degradation prior to conversion by incubating DNA conversion reagent (e.g. bisulfite reagent) directly with a nucleic acid containing sample without requiring prior nucleic acid purification from the sample and without requiring prior nucleic acid denaturation at elevated temperatures of 98° C. and, b) optimizing bisulfite removal by controlling the pumping rate flow of the bisulfite treated sample over an extraction membrane inside an automated system. In some embodiments, a cartridge for the detection of methylated DNA is provided.

The present invention is based on the discovery of sensitive and specific methylation detection by a) controlling excessive DNA degradation prior to conversion by incubating DNA conversion reagent (e.g. bisulfite reagent) directly with a nucleic acid containing sample without requiring prior nucleic acid purification from the sample and without requiring prior nucleic acid denaturation at elevated temperatures of 98° C. and, b) optimizing bisulfite removal by controlling the pumping rate flow of the bisulfite treated sample over an extraction membrane inside an automated system. In some embodiments, a cartridge for the detection of methylated DNA is provided.

Provided herein are methods, compositions, and kits for forming amplification products. In various embodiments provided herein, transposomes comprising transposases are used in forming tagged polynucleotides for downstream amplification and polynucleotide processing steps.

Provided herein are methods, compositions, and kits for forming amplification products. In various embodiments provided herein, transposomes comprising transposases are used in forming tagged polynucleotides for downstream amplification and polynucleotide processing steps.