C12Q2521/514

High throughput measurement of DNA base lesion repair capacity

The invention pertains to methods for measuring DNA repair capacity of a test solution, for example, a cell lysate. The method can comprise incubating the test solution with a damaged DNA molecule and measuring the damaged DNA remaining at the end of the incubation. The damaged DNA used in the method comprises a marker attached to a first strand of the damaged DNA and a quencher attached to a second strand of the damaged DNA, characterized in that the quencher inhibits the release of a detectable signal from the marker when damaged DNA is in a double stranded form and the marker emits the detectable signal when at least a portion of the damaged DNA is in a single stranded form. The invention also provides screening assays for identifying one or more compounds from a plurality of compounds as an inhibitor of DNA repair mechanism.

DETECTION OF MUTATIONS REGARDING ONE OR MORE DEOXYRIBONUCLEIC ACID SEQUENCES USING DETERMINISTIC LATERAL DISPLACEMENT ARRAYS

Techniques regarding screening for mutations using nanoscale deterministic arrays are provided. For example, one or more embodiments described herein can comprise a method, which can comprise cleaving a deoxyribonucleic acid segment hybridized with a molecular probe to form a sample fluid. The cleaving can occur at a first end and a second end of the molecular probe. Also, the cleaving can comprise a cleaving agent that targets base pair mismatches. The method can also comprise supplying the sample fluid to a nanoscale deterministic lateral displacement array to screen for a single nucleotide polymorphism.

DETECTION OF MUTATIONS REGARDING ONE OR MORE DEOXYRIBONUCLEIC ACID SEQUENCES USING DETERMINISTIC LATERAL DISPLACEMENT ARRAYS

Techniques regarding screening for mutations using nanoscale deterministic arrays are provided. For example, one or more embodiments described herein can comprise a method, which can comprise cleaving a deoxyribonucleic acid segment hybridized with a molecular probe to form a sample fluid. The cleaving can occur at a first end and a second end of the molecular probe. Also, the cleaving can comprise a cleaving agent that targets base pair mismatches. The method can also comprise supplying the sample fluid to a nanoscale deterministic lateral displacement array to screen for a single nucleotide polymorphism.

Materials and methods for the synthesis of error-minimized nucleic acid molecules

The present invention provides materials and methods useful for error correction of nucleic acid molecules. In one embodiment of the invention, a first plurality of double-stranded nucleic acid molecules having a nucleotide mismatch are fragmented by exposure to a molecule having unidirectional mismatch endonuclease activity. The nucleic acid molecules are cut at the mismatch site or near the mismatch site, leaving a double-stranded nucleic acid molecule having a mismatch at the end or near end of the molecule. The nucleic acid molecule is then exposed to a molecule having unidirectional exonuclease activity to remove the mismatched nucleotide. The missing nucleotides can then be filled in by the action of, e.g., a molecule having DNA polymerase activity. The result is double-stranded nucleic acid molecules with a decreased frequency of nucleotide mismatches. Also provided are novel nucleic acid sequences encoding mismatch endonucleases, polypeptides encoded thereby, as well as nucleic acid constructs, transgenic cells, and various compositions thereof.

Materials and methods for the synthesis of error-minimized nucleic acid molecules

The present invention provides materials and methods useful for error correction of nucleic acid molecules. In one embodiment of the invention, a first plurality of double-stranded nucleic acid molecules having a nucleotide mismatch are fragmented by exposure to a molecule having unidirectional mismatch endonuclease activity. The nucleic acid molecules are cut at the mismatch site or near the mismatch site, leaving a double-stranded nucleic acid molecule having a mismatch at the end or near end of the molecule. The nucleic acid molecule is then exposed to a molecule having unidirectional exonuclease activity to remove the mismatched nucleotide. The missing nucleotides can then be filled in by the action of, e.g., a molecule having DNA polymerase activity. The result is double-stranded nucleic acid molecules with a decreased frequency of nucleotide mismatches. Also provided are novel nucleic acid sequences encoding mismatch endonucleases, polypeptides encoded thereby, as well as nucleic acid constructs, transgenic cells, and various compositions thereof.

IMPROVED NUCLEIC ACID TARGET ENRICHMENT AND RELATED METHODS
20220136042 · 2022-05-05 ·

The invention is an improved method of target enrichment and target depletion where probe binding is facilitated by the FANCA protein. Improved workflows for next regeneration sequencing and related methods involving nucleic acid probe hybridization are improved by the use of FANCA protein.

IMPROVED NUCLEIC ACID TARGET ENRICHMENT AND RELATED METHODS
20220136042 · 2022-05-05 ·

The invention is an improved method of target enrichment and target depletion where probe binding is facilitated by the FANCA protein. Improved workflows for next regeneration sequencing and related methods involving nucleic acid probe hybridization are improved by the use of FANCA protein.

Mapping the location, type and strand of damaged and/or mismatched nucleotides in double-stranded DNA

Providing herein, among other things, is a method comprising incubating a double-stranded nucleic acid having a nick with a nick translating activity, a ligase, and a nucleotide mix comprising at least one modified nucleotide, to generate a product comprising a patch of a newly synthesized strand of a duplex nucleic acid containing a plurality of modified nucleoside monophosphates that are at or adjacent to the site of the nick. In some embodiments, the method may be used to map damaged nucleoside monophosphates in a nucleic acid. Compositions and kits for use in performing the method are also provided.

Mapping the location, type and strand of damaged and/or mismatched nucleotides in double-stranded DNA

Providing herein, among other things, is a method comprising incubating a double-stranded nucleic acid having a nick with a nick translating activity, a ligase, and a nucleotide mix comprising at least one modified nucleotide, to generate a product comprising a patch of a newly synthesized strand of a duplex nucleic acid containing a plurality of modified nucleoside monophosphates that are at or adjacent to the site of the nick. In some embodiments, the method may be used to map damaged nucleoside monophosphates in a nucleic acid. Compositions and kits for use in performing the method are also provided.

Detection of mutations regarding one or more deoxyribonucleic acid sequences using deterministic lateral displacement arrays

Techniques regarding screening for mutations using nanoscale deterministic arrays are provided. For example, one or more embodiments described herein can comprise a method, which can comprise cleaving a deoxyribonucleic acid segment hybridized with a molecular probe to form a sample fluid. The cleaving can occur at a first end and a second end of the molecular probe. Also, the cleaving can comprise a cleaving agent that targets base pair mismatches. The method can also comprise supplying the sample fluid to a nanoscale deterministic lateral displacement array to screen for a single nucleotide polymorphism.