Abstract: A method of preparing a sample may include depositing an aqueous solution comprising copies of a primer into a layer of hydrophobic liquid on a substrate with a thermal inkjet device. A sample may include: a substrate; a layer of hydrophobic liquid on the substrate, the layer of hydrophobic liquid comprising a plurality of droplets of aqueous solution distributed in the layer, wherein the plurality of droplets contain: primers; a polymerase enzyme; deoxynucleotide triphosphates (dNTPs); and a target sequence for replication; and a cover, the cover contacting and covering the layer of hydrophobic liquid.

Abstract: Disclosed is a system and method for production of DNA particles and use thereof. The DNA particles can be produced by amplification of nucleic acid molecule(s). Alternatively, DNA particles can be prepared by condensing multiple DNA molecules. The DNA condensation into a particle is mainly triggered by pyrophosphate and positively charged cations (e.g. magnesium). DNA particles can be applied for numerous biological applications but not limited to directed evolution, proteomics, drug delivery and imaging. DNA particles can be used to synthesize proteins using in vitro transcription/translation reaction.

Abstract: The present disclosure provides methods, compositions and kits as well as systems for manipulating nucleic acids, including implementing isothermal amplification, such as recombinase-polymerase amplification (RPA), of a nucleic acid template using a pre-seeded solid support. Provided are rapid and efficient methods for generating template nucleic acid molecules comprising specific nucleotide sequence bound to solid support. Such methods can be used, for example, in manipulating nucleic acids in preparation for analysis methods that utilize monoclonal populations of nucleic acids.

Abstract: The present application provides a method for genotyping M. tuberculosis, comprising obtaining amplifying and obtaining a first DNA fragment from a DNA sample by using one or more primer sets selected from the group consisting of primer sets 1 to 25 (SEQ ID Nos. 1 to 50); amplifying and obtaining a second DNA fragment from the obtained first DNA fragment by using one or more extension primers selected from the group consisting of SEQ ID Nos. 51 to 75; and detecting the second DNA fragment by using mass spectrometry, particularly by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS).

Abstract: A primer set includes a terminal primer A including, in its 3?-terminal part, a nucleotide sequence that hybridizes to a 3?-terminal part of a complementary sequence of a first target nucleic acid sequence, a k-th double-headed primer including two polynucleotides linked at their 5? terminal sides, wherein one of the two polynucleotides includes, in its 3?-terminal part, a nucleotide sequence that hybridizes to a 3?-terminal part of a nucleotide sequence of a k-th target nucleic acid, and the other polynucleotide includes, in its 3?-terminal part, a nucleotide sequence that hybridizes to a 3?-terminal part of a complementary sequence of a (k+1)th target nucleic acid, and a terminal primer B including, in its 3?-terminal part, a nucleotide sequence that hybridizes to a 3?-terminal part of a nucleotide sequence of a N-th target nucleic acid.

Abstract: Minimal-copy-ratio of templates is a problem in detecting early stage cancer where minimal copies of somatic cancer-specific mutations are targeted in the presence of large copies of wildtype genome DNA, commonly a 1/10,000 or even less minimal-copy-ratio between the mutant target and wildtype control templates. To overcome this problem, delayed pyrophosphorolysis activated polymerization (delayed-PAP) was developed which can delay product accumulation of the wildtype control to a much later time or cycle by up to 15 cycles or by 30,000 folds. In the multiplex format, delayed-PAP is particularly useful to amplify not only the wildtype control but also mutant target templates accurately and consistently in the minimal-copy-ratio situation.

Abstract: Provided is a nucleic acid isothermal self-amplification method comprising, adding suitable palindrome complementary sequences at both ends of a target template to form a stem-loop structure spontaneously, and providing reagents and conditions as needed to perform self-amplification. The method does not require addition of additional amplification primers. The reagent comprises a DNA polymerase having a strand displacement activity. The method does not rely on exogenous amplification primers for amplification, has a constant amplification temperature without a complex temperature control equipment, and achieves rapid amplification. The amplification product is a long single-stranded DNA of a continuous complementary sequence and can be applied to special occasions. In addition, the amplification has no GC bias.

Abstract: Methods and compositions are provided for quantifying the number of biological input molecules of one or more target genes of interest in a PCR. The method comprises steps for amplifying multiple synthetic templates and high throughput sequencing. The method further comprises steps for achieving equal sequencing coverage between targets that do not occur in equal ratios, while mitigating the need for costly deep sequencing methods.

Abstract: Customization of a universal reagent cartridge with a lyophilized target-specific PCR component reagent is disclosed. The universal reagent cartridge includes multiple chambers populated with predetermined non-target-specific reagents therein. A custom reagent storage device that stores a lyophilized, target-specific reagent is provided and is configured to be end user insertable into the universal reagent cartridge to customize the universal reagent cartridge with the target-specific reagent.

Abstract: A method of digital quantification of a species in an EWOD device includes inputting a sample volume and a diluent volume into the EWOD device; performing an electrowetting operation to generate a first sample droplet from the sample volume; performing an amplification process on the first sample droplet and measuring a turn-on value for the sample droplet; comparing the measured turn-on value to a target turn-on value for digital quantification; calculating a dilution factor based on the comparison of the measured and target turn-on values; performing an electrowetting operation to extract a second sample droplet from the sample volume; performing an electrowetting operation to dilute the second sample droplet with the diluent volume by the dilution factor to form a diluted second sample droplet; and performing a digital quantification on the diluted second sample droplet to quantify an initial concentration of the species in the sample volume.

Abstract: Provided herein are, inter alia, methods and compositions to detect, monitor and treat cancer, wherein the cancer includes amplified extrachromosomal oncogenes. The methods are useful for personalized treatment and exploit differential expression of amplified extrachromosomal oncogenes in cancer cells versus healthy cells.

Abstract: The present invention relates to a method and a composition for detecting a target nucleic acid sequence using a cleaved complementary tag fragment. Specifically, the present invention relates to a method for linking a complementary tag sequence to a PCR primer so that a tagging can be produced by a restriction enzyme during a PCR reaction, diversifying the complementary tag sequence to be linked to each primer by utilizing factors such as length and nucleic acid combination, etc., and distinguishing the target sequence using the same.

Abstract: A method includes coupling a molecular diagnostic test device to a power source. A biological sample is conveyed into a sample preparation module. The device is then actuated by only a single action to cause the device to perform the following functions without further user action. First, the device heats the sample via a heater of the sample preparation module to lyse a portion of the sample. Second, the device conveys the lysed sample to an amplification module and heats the sample within a reaction volume of the amplification module to amplify a nucleic acid thereby producing an output solution containing a target amplicon. The device then reacts, within a detection module, each of (i) the output solution and (ii) a reagent formulated to produce a signal that indicates a presence of the target amplicon within the output solution. A result associated with the signal is then read.

Abstract: The present disclosure provides a nucleic acid amplification method. The method comprises forming an enrichment culture by contacting a sample with a nutrient medium having a formulation that does not include a phosphate buffer component; holding the enrichment culture for a period of time at a temperature that facilitates growth of a target microorganism; after holding the enrichment culture, forming an aqueous composition by mixing a first volume of the enrichment culture with a second volume of a lysis buffer; contacting the aqueous composition with an effective amount of a water-insoluble material that sequesters a substance that interferes with a polymerase-mediated nucleic acid amplification reaction; subjecting the aqueous composition to a thermal lysis process; and, after subjecting the aqueous composition to the thermal lysis process, subjecting a portion of the aqueous composition to a nucleic acid amplification process. A composition for a lysis buffer is also disclosed.

Abstract: A method includes coupling a molecular diagnostic test device to a power source. A biological sample is conveyed into a sample preparation module. The device is then actuated by only a single action to cause the device to perform the following functions without further user action. First, the device heats the sample via a heater of the sample preparation module to lyse a portion of the sample. Second, the device conveys the lysed sample to an amplification module and heats the sample within a reaction volume of the amplification module to amplify a nucleic acid thereby producing an output solution containing a target amplicon. The device then reacts, within a detection module, each of (i) the output solution and (ii) a reagent formulated to produce a signal that indicates a presence of the target amplicon within the output solution. A result associated with the signal is then read.

Abstract: The invention is a novel method of generating a library of circular single stranded nucleic acid molecules by utilizing circular capture molecules. The method is not limited by size of target nucleic acid molecules and can potentially accommodate very long molecules. The method finds application in nucleic acid sequencing, e.g., nanopore sequencing where unlimited-length templates can be read.

Abstract: A multiplex slide plate device is provided, including a slide plate, a sacrificial layer, and a housing. The slide plate has reaction vessels arranged in an array. The sacrificial layer has a microfluidic channel, which has an injection channel, a main channel, and a distal channel connected to each other. The housing is used to accommodate the slide plate and the sacrificial layer, and is composed of a cover and a tray, and the cover has an injection hole, an exhaust hole, and a storage tank. A sample solution and an oil are injected from the injection hole into the injection channel, wherein the sample solution is pushed by the oil. The sample solution loads into each of the reaction vessels while flowing through the main channel. Excess waste liquid flows from the exhaust hole into the storage tank, and is covered by the oil and cannot reflow.

Abstract: Provided herein is technology for neoplasia screening, and particularly, but not exclusively, to methods, compositions, and related uses for detecting the presence of cancer, in particular, colorectal cancer.

Abstract: One aspect of the invention is a method for amplifying alpha globin genes HBA1, HBA2 and HBA12 in a single PCR tube to determine an HBA genotype of a subject. This method employs five primers selected to accurate and sensitively identify the HBA1, HBA2, and HBA12, a gene found at a higher frequency in citizens of Saudi Arabia, by accurately annealing to nucleic acids in a biological sample and simultaneously amplifying sequences encoding the alpha globin genes. This invention includes a procedure and required reagents for the amplification of alpha globin genes in a single PCR tube.

Abstract: The present invention refers to an in vitro method for screening for subjects at risk of developing pancreatic cancer or intraductal papillary mucinous neoplasm of the pancreas (IPMN) comprising: (a) measuring the expression pattern or level of at least hsa-miR-33a*, or of at least hsa-miR-320a, or of at least hsa-let-7e, or of at least hsa-let-7f, or of at least hsa-miR-1257, or of at least hsa-miR-1304, or of at least hsa-miR-151b, or of at least hsa-miR-3120-3p, or of at least hsa-miR-3133, or of at least hsa-miR-3714, or of at least hsa-miR-4468, or of at least hsa-miR-4639-5p, or of at least hsa-miR-4713-5p, or of at least hsa-miR-4714-5p, or of at least hsa-miR-4770, or of at least hsa-miR-548d-3p, or of at least hsa-miR-761, obtained from an isolated biological sample of the subjects to be screened; and (b) comparing said expression pattern or level of at least hsa-miR-33a*, or of at least hsa-miR-320a, or of at least hsa-let-7e, or of at least hsa-let-7f, or of at least hsa-miR-1257, or of at least hs