Method for Library Preparation in Next Generation Sequencing by Enzymatic DNA Fragmentation

    公开(公告)号:US20240076803A1

    公开(公告)日:2024-03-07

    申请号:US18274541

    申请日:2022-01-28

    发明人: Matthias Wahl

    IPC分类号: C40B50/06

    CPC分类号: C40B50/06

    摘要: The invention is directed to a method for obtaining a nucleic acid library of a sample comprising polynucleotides comprising the steps a. multiplying the polynucleotides by a polymerase b. fragmenting the multiplied polynucleotides by creating nicks c. coupling an oligonucleotide sequence to the nicks to create the target library wherein step a) is performed by providing A, T, G, C and U nucleotides wherein the molar ratio of T and U is between 200:1 and 5:1 and step b) is performed by excision of the U nucleotides. characterized in that after step b), the nicks are provided with a polymerase exhibiting 5′ #3′ exonuclease activity, thereby filling in the 3′ recessing ends and removing the 5′ overhangs of the nicks.

    CONJUGATES HAVING  AN ENZYMMATICALLY RELEASABLE DETECTION MOIETY AND A BARCODE MOIETY

    公开(公告)号:US20230324399A1

    公开(公告)日:2023-10-12

    申请号:US18024603

    申请日:2020-09-07

    发明人: Andreas Bosio

    IPC分类号: G01N33/58 C40B30/04

    摘要: The invention is directed to a conjugate having the general formula (I): Xn—P—YmBo (I), with X is an detection moiety, P is a spacer unit, Y an antigen recognizing moiety, B an oligonucleotide comprising 2 to 300 nucleotide residues and n, m, o are independent integers between 1 and 100 wherein P and B are covalently bound to Y and X is covalently bound to P and wherein X is erasable. Further, the invention is directed to a library of such conjugates and a method of detecting target cells utilizing the conjugates or the library of conjugates.

    Water-soluble M-conjugated fluorescent 1,1-binaphthyl-based tandem polymers

    公开(公告)号:US20230228746A1

    公开(公告)日:2023-07-20

    申请号:US18097244

    申请日:2023-01-15

    IPC分类号: G01N33/533 C08G61/12

    摘要: The invention is directed to a conjugate having the general formula (I)






    Wherein AR, MU and MU* are repeating units of a polymer and MU and MU* are polymer modifying units or band gap modifying units which are evenly or randomly distributed along the polymer main chain,
    G1 and G2 stand for hydrogen, halogen or an antigen recognizing moiety, with the provision that at least one of G1 or G2 is an antigen recognizing moiety,
    a is 10 to 100 mol %,
    b is 0 to 90 mol %
    c is 0.1 to 90 mol %
    d is 1 to 10 000; with the provisio that a+b+c=100 mol %
    characterized in that AR is connected in the polymer chain via the 2,2′ or 3,3′ or 4,4′ or 5,5′ or 6,6′ or 7,7′ or 8,8′ positions according to general formula (II)










    Wherein the remaining positions 2,2′; 3,3′; 4,4′; 5,5′; 6,6′; 7,7′ and 8,8′ are substituted with same or different residues selected from the group consisting of H, SO2CF3, SO2Ra, CF3, CCl3, CN, SO3H, NO2, NRaRbRc+, CHO, CORa, CO2Ra, COCl, CONRaRb, F, Cl, Br, I, Ra, ORa, SRa, OCORa, NRaRb, NHCORa, CCRa, aryl-, heteroaryl-, C6H4ORa or C6H4NRaRb, with Ra-c independently hydrogen, alkyl-, alkenyl-, alkinyl-, heteroalkyl-, aryl-, heteroaryl-, cycloalkyl-, alkylcycloalkyl-, heteroalkylcycloalkyl-, heterocycloalkyl-, aralkyl- or a heteroaralkyl residue or (CH2)x(OCH2CH2)yO(CH2)zCH3, wherein x is an integer from 0 to 20; y is an integer from 0 to 50 and z is an integer from 0 to 20.

    Perestaltic pump and device for isolating cells from biological tissue

    公开(公告)号:US11639717B2

    公开(公告)日:2023-05-02

    申请号:US16833694

    申请日:2020-03-30

    IPC分类号: F04B43/14 A01N1/02 A61M60/279

    摘要: The invention is directed to a peristaltic pump (10), comprising: a flexible membrane (1) forming a at least one bladder (2) against a support, wherein each bladder is provided with one input orifice (4) which admits a fluid to the bladder (2) and one outlet orifice (5) which releases the fluid from the bladder (2); and at least one roller bearing (6) is configured to rotate about an axis (7) and to apply a compressive force against the at least one bladder (2). The peristaltic pump may be in fluid communication with a tapered jet (11).

    LABELED PROBES WITH DIFFERENTIALLY CLEAVABLE LINKERS AND THEIR USE IN DE-CODING DNA AND RNA MOLECULES

    公开(公告)号:US20230101711A1

    公开(公告)日:2023-03-30

    申请号:US17952980

    申请日:2022-09-26

    IPC分类号: C12Q1/6874 C12N15/10

    摘要: The invention is directed to a method for detecting RNA, DNA or protein target sequences by a) Hybridizing a library of probes having the general formula (I) P—(CL-D)x  (I)  With P: probes having at least 10 nucleotides or amino acids CL: cleavable linker D: fluorescent dye X: integer between 1 and 5  to RNA, DNA or protein target sequences wherein the library comprises probes P having different sequences of nucleotides or amino acids and cleavable linkers CL of different groups which are cleavable with different means b) Removing unhybridized probes and detecting the hybridized probes via the fluorophores D by a first image c) Cleaving sequentially by different means each group of chemical linkers CL from the hybridized probes; removing the thus cleaved fluorophores D and detecting the remaining hybridized probes via their fluorophores D by a second image d) Detecting the removed fluorophores D by comparing the first and second image. e) Obtaining a part of the sequence information of the target sequences via the sequence information of the probes P associated with the removed fluorophores D f) Repeating step c) until all groups of chemical linkers CL are cleaved.