Described herein are synthetic methods for producing sequence-specific RNA oligonucleotides that eliminate impurities produced in prior art methods. In one aspect, an end-protected capture DNA complementary to a portion of the product RNA is employed. In another aspect, the template DNA is covalently or noncovalently linked to the RNA polymerase, either directly or through the use of a nontemplate DNA. In a third aspect, a flow chamber is employed. All of the methods can be used in combination.

Described herein are synthetic methods for producing sequence-specific RNA oligonucleotides that eliminate impurities produced in prior art methods. In one aspect, an end-protected capture DNA complementary to a portion of the product RNA is employed. In another aspect, the template DNA is covalently or noncovalently linked to the RNA polymerase, either directly or through the use of a nontemplate DNA. In a third aspect, a flow chamber is employed. All of the methods can be used in combination.

Described herein are synthetic methods for producing sequence-specific RNA oligonucleotides that eliminate impurities produced in prior art methods. In one aspect, an end-protected capture DNA complementary to a portion of the product RNA is employed. In another aspect, the template DNA is covalently or noncovalently linked to the RNA polymerase, either directly or through the use of a nontemplate DNA. In a third aspect, a flow chamber is employed. All of the methods can be used in combination.

The present invention relates to a detection method for detecting a target RNA contained in a sample with high sensitivity by using nicking/extension chain reaction system-based isothermal nucleic acid amplification (NESBA) that uses activity of a cleavage enzyme and a DNA polymerase. The NESBA of the present invention is a new concept isothermal target RNA detection method that realizes higher amplification efficiency than the existing NASBA technology and is deemed to be utilizable as a new concept diagnosis technology that can replace conventional target RNA detection technologies.

The present invention relates to a detection method for detecting a target RNA contained in a sample with high sensitivity by using nicking/extension chain reaction system-based isothermal nucleic acid amplification (NESBA) that uses activity of a cleavage enzyme and a DNA polymerase. The NESBA of the present invention is a new concept isothermal target RNA detection method that realizes higher amplification efficiency than the existing NASBA technology and is deemed to be utilizable as a new concept diagnosis technology that can replace conventional target RNA detection technologies.

The present disclosure belongs to the technical field of genetic modification of an enzyme preparation and particularly relates to a preparation method of a high-stability superoxide dismutase with a transmembrane capability. The method includes the following steps: extracting mRNA from Geobacillus stearothermophilus, synthesizing cDNA by a reverse transcription method, amplifying a large number of coding regions of the cDNA by designing a specific primer, ligating the coding regions to an E. coli expression vector, and transforming the coding regions into engineering bacteria BL21 (DE3). A point mutation technology is used to enhance stability of the superoxide dismutase and a flexible polypeptide linker GGGSGGGS (SEQ ID NO: 11) is designed, such that a soluble fusion expression of a transmembrane peptide YGRKKRRQRRR (SEQ ID NO: 10) and the superoxide dismutase is successfully realized.

The present disclosure belongs to the technical field of genetic modification of an enzyme preparation and particularly relates to a preparation method of a high-stability superoxide dismutase with a transmembrane capability. The method includes the following steps: extracting mRNA from Geobacillus stearothermophilus, synthesizing cDNA by a reverse transcription method, amplifying a large number of coding regions of the cDNA by designing a specific primer, ligating the coding regions to an E. coli expression vector, and transforming the coding regions into engineering bacteria BL21 (DE3). A point mutation technology is used to enhance stability of the superoxide dismutase and a flexible polypeptide linker GGGSGGGS (SEQ ID NO: 11) is designed, such that a soluble fusion expression of a transmembrane peptide YGRKKRRQRRR (SEQ ID NO: 10) and the superoxide dismutase is successfully realized.

The present invention relates to assays, including amplification assays, conducted in the presence of modulators. These assays can be used to detect the presence of particular nucleic acid sequences. In particular, these assays can allow for genotyping or other genetic analysis.

The methods, compositions, and kits of the disclosure provide a novel approach for a whole genome, unbiased DNA analysis method that can be performed on limited amounts of DNA. can be used to analyze DNA to determine its modification status. Aspects of the disclosure relate to a method for amplifying bisulfite-treated deoxyribonucleic acid (DNA) molecules comprising: (a) ligating an adaptor to the DNA molecules, wherein the adaptor comprises a RNA polymerase promoter comprising bisulfite-protected cytosines; (b) treating the ligated DNA molecules with bisulfite; (c) hybridizing the bisulfite-treated DNA molecules with a primer; (d) extending the hybridized primer to make double stranded DNA; and (e) in vitro transcribing the double-stranded DNA to make RNA.

The methods, compositions, and kits of the disclosure provide a novel approach for a whole genome, unbiased DNA analysis method that can be performed on limited amounts of DNA. can be used to analyze DNA to determine its modification status. Aspects of the disclosure relate to a method for amplifying bisulfite-treated deoxyribonucleic acid (DNA) molecules comprising: (a) ligating an adaptor to the DNA molecules, wherein the adaptor comprises a RNA polymerase promoter comprising bisulfite-protected cytosines; (b) treating the ligated DNA molecules with bisulfite; (c) hybridizing the bisulfite-treated DNA molecules with a primer; (d) extending the hybridized primer to make double stranded DNA; and (e) in vitro transcribing the double-stranded DNA to make RNA.