LINEAR DNA WITH ENHANCED RESISTANCE AGAINST EXONUCLEASES AND METHODS FOR THE PRODUCTION THEREOF

    公开(公告)号:US20240360488A1

    公开(公告)日:2024-10-31

    申请号:US18429780

    申请日:2024-02-01

    Abstract: Methods for producing a linear deoxyribonucleic acid (DNA) product with enhanced resistance to nuclease digestion are provided. The methods comprise, (a) digesting a double-stranded DNA molecule with an endonuclease that cleaves an endonuclease target sequence to generate a digested double-stranded DNA molecule, wherein the digested double-stranded DNA molecule comprises a linear double-stranded region, and a truncated protelomerase sequence at a first end, wherein the truncated protelomerase target sequence is non-functional; (b) appending a first adaptor molecule to the first end of the digested double-stranded DNA molecule and appending a second adaptor molecule to the second end of the digested double-stranded DNA molecule to generate a precursor double-stranded DNA molecule, wherein the first adaptor molecule comprises a truncated protelomerase target sequence that forms a first functional protelomerase target sequence with the truncated protelomerase sequence at the first end of the digested double-stranded DNA molecule; and (c) incubating the precursor double-stranded DNA molecule with a protelomerase to generate the linear DNA product, wherein the protelomerase closes the first end of the precursor double-stranded DNA molecule at the first functional protelomerase target sequence.

    Method for Producing Circular DNA
    4.
    发明公开

    公开(公告)号:US20240352495A1

    公开(公告)日:2024-10-24

    申请号:US18685259

    申请日:2022-09-13

    Abstract: Provided is a method for producing circular DNA in which a region that is sandwiched by a region Ha and a region Hb in circular double-stranded DNA is substituted with the entirety or a portion of a linear-DNA fragment, wherein the region Hb is located downstream of the region Ha in the circular double-stranded DNA; the linear-DNA fragment is single-stranded or double-stranded linear DNA that has a homologous region that corresponds to the region Ha and a homologous region that corresponds to the region Hb, the latter homologous region being positioned downstream of the former homologous region; and the method comprising: preparing a reaction solution that contains the circular double-stranded DNA, the linear-DNA fragment, and a protein that has RecA-family-recombinase activity, and performing homologous recombination reaction by incubating the reaction solution for a predetermined period of time, thereby producing circular DNA in which the region that is from the region Ha to the region Hb in the circular double-stranded DNA is substituted with the region that is from the homologous region that corresponds to the region Ha to the homologous region that corresponds to the region Hb in the linear-DNA fragment.

    COMPOSITIONS AND METHODS FOR NUCLEIC ACID AMPLIFICATION

    公开(公告)号:US20240254546A1

    公开(公告)日:2024-08-01

    申请号:US18487416

    申请日:2023-10-16

    Applicant: AMPLIWISE INC.

    Inventor: Kai Wu Mindy Su Xing Su

    CPC classification number: C12Q1/6853 C12P19/34 C12Q1/6827 C12Q1/686

    Abstract: An embodiment relates to system comprising: (i) a reaction mixture comprises: a) a target nucleic acid molecule; b) a forward primer complementary to a strand of the target nucleic acid molecule, the forward primer comprises a first molecular moiety at a 3′ end, wherein the first molecular moiety is non-complementary to the strand of the target nucleic acid molecule; c) a reverse primer complementary to a complementary sequence of the strand of the target nucleic acid molecule, the reverse primer comprises a second molecular moiety at a 3′ end, wherein the second molecular moiety is non-complementary to the complementary sequence of the strand of the target nucleic acid molecule; d) a polymerase with 3′-5′ exonuclease activity; and (ii a device suitable of detecting an amplification products.

Patent Agency Ranking