Patent classifications
C12Q2527/101
METHODS AND COMPOSITIONS RELATING TO HOT-START, ONE-STEP, REVERSE TRANSCRIPTION-COUPLED PCR
Methods of amplifying an RNA template according to aspects of the present disclosure include: providing a composition including a recombinant thermostable DNA polymerase including SEQ ID NO:1, or a variant thereof having at least 99% identity to SEQ ID NO:1; and a reaction buffer, the reaction buffer including 10-30 mM Tris-HCl, pH 8.5-9.0; 20-40 mM KCl; 5-15 mM (NH.sub.4).sub.2SO.sub.4; 1.5-3.5 mM Mn.sup.2+; 0.01-0.20% (w/v) gelatin and/or serum albumin; 0.05-0.15% (w/v) of a nonionic detergent; with the proviso that no more than 0.1 mM of Mg.sup.2+ is present in the composition; and, optionally, 0.01-0.1 mM of a chelating agent is present in the composition.
METHODS AND COMPOSITIONS RELATING TO HOT-START, ONE-STEP, REVERSE TRANSCRIPTION-COUPLED PCR
Methods of amplifying an RNA template according to aspects of the present disclosure include: providing a composition including a recombinant thermostable DNA polymerase including SEQ ID NO:1, or a variant thereof having at least 99% identity to SEQ ID NO:1; and a reaction buffer, the reaction buffer including 10-30 mM Tris-HCl, pH 8.5-9.0; 20-40 mM KCl; 5-15 mM (NH.sub.4).sub.2SO.sub.4; 1.5-3.5 mM Mn.sup.2+; 0.01-0.20% (w/v) gelatin and/or serum albumin; 0.05-0.15% (w/v) of a nonionic detergent; with the proviso that no more than 0.1 mM of Mg.sup.2+ is present in the composition; and, optionally, 0.01-0.1 mM of a chelating agent is present in the composition.
PROCESSES FOR PURIFYING DOWNSTREAM PRODUCTS OF IN VITRO TRANSCRIPTION
Provided herein, in some embodiments, are methods of purifying low-salt RNA compositions using denaturing oligo-dT chromatography.
PROCESSES FOR PURIFYING DOWNSTREAM PRODUCTS OF IN VITRO TRANSCRIPTION
Provided herein, in some embodiments, are methods of purifying low-salt RNA compositions using denaturing oligo-dT chromatography.
CAP GUIDES AND METHODS OF USE THEREOF FOR RNA MAPPING
The present disclosure relates, in some embodiments, to isolated nucleic acids (also referred to as cap guides) and methods of use thereof for RNA mapping. The disclosure is based, in part, on guide RNAs that bind to a position that is at least 7 nucleotides downstream of the first nucleotide of an mRNA molecule.
CAP GUIDES AND METHODS OF USE THEREOF FOR RNA MAPPING
The present disclosure relates, in some embodiments, to isolated nucleic acids (also referred to as cap guides) and methods of use thereof for RNA mapping. The disclosure is based, in part, on guide RNAs that bind to a position that is at least 7 nucleotides downstream of the first nucleotide of an mRNA molecule.
RNASE H2 MUTANTS THAT REDUCE PRIMER DIMERS AND OFF-TARGET AMPLIFICATION IN RHPCR-BASED AMPLICON SEQUENCING WITH HIGH-FIDELITY DNA POLYMERASES
The present invention pertains to hybrid RNase H2 proteins that include fragments of amino acid sequences from Pyrococcus abyssi (P.a.), Thermococcus kodakarensis (T.kod), and Pyrococcus furiosus organisms, as well as methods of using the same to improve mismatch discrimination and activity in a high-fidelity DNA polymerase buffer.
RNASE H2 MUTANTS THAT REDUCE PRIMER DIMERS AND OFF-TARGET AMPLIFICATION IN RHPCR-BASED AMPLICON SEQUENCING WITH HIGH-FIDELITY DNA POLYMERASES
The present invention pertains to hybrid RNase H2 proteins that include fragments of amino acid sequences from Pyrococcus abyssi (P.a.), Thermococcus kodakarensis (T.kod), and Pyrococcus furiosus organisms, as well as methods of using the same to improve mismatch discrimination and activity in a high-fidelity DNA polymerase buffer.
Method of detecting genetic material in a biological sample and a device for its implementation
The object of the invention is a method of detecting genetic material in a biological sample in which the biological sample is loaded into the reaction cartridge (6) and then the reaction cartridge (6) is placed in the control device, the collected biological sample is taken to the isolation chamber (7), isolation of biological material from the tested sample by heating the isolation chamber (7), the isolated genetic material is moved into a plurality of reaction chambers (8.1, 8.2, 8.3, 8.4), genetic material is amplified by heating the reaction chambers (8.1, 8.2, 8.3, 8.4), lyophilized reagents for genetic material amplification together with lyophilized fluorescent tag intercalating with genetic material are present in the reaction chambers (8.1, 8.2, 8.3, 8.4), and signal detection from fluorescent tags is carried out along with the genetic material amplification stage.
Method of detecting genetic material in a biological sample and a device for its implementation
The object of the invention is a method of detecting genetic material in a biological sample in which the biological sample is loaded into the reaction cartridge (6) and then the reaction cartridge (6) is placed in the control device, the collected biological sample is taken to the isolation chamber (7), isolation of biological material from the tested sample by heating the isolation chamber (7), the isolated genetic material is moved into a plurality of reaction chambers (8.1, 8.2, 8.3, 8.4), genetic material is amplified by heating the reaction chambers (8.1, 8.2, 8.3, 8.4), lyophilized reagents for genetic material amplification together with lyophilized fluorescent tag intercalating with genetic material are present in the reaction chambers (8.1, 8.2, 8.3, 8.4), and signal detection from fluorescent tags is carried out along with the genetic material amplification stage.