IP Library › Granted Patent US 12,209,276
Granted Patent B2
US 12,209,276 · App. 18/494,603 · Granted Jan 28, 2025

Hybridization compositions and methods

Inventors: Charles M. Hansen (Horsholm, DK); Steen Hauge Matthiesen (Hillerød, DK); Kenneth H. Petersen (Smørum, DK); Tim Svenstrup Poulsen (Horsholm, DK)
Assignee: Agilent Technologies, Inc.
C12Q1/6832C12Q1/6841Y10T436/143333
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Quick Facts
Patent No.
US 12,209,276
App. No.
18/494,603
Granted
Jan 28, 2025
Kind
B2
Abstract

The invention provides methods and compositions for hybridizing at least one molecule to a target. The invention may, for example, eliminate the use of, or reduce the dependence on formamide in hybridization. Compositions for use in the invention include an aqueous composition comprising at least one nucleic acid sequence and at least one polar aprotic solvent in an amount effective to denature double-stranded nucleotide sequences.

Claims (20)

1. A hybridization composition comprising a mixture of at least one nucleic acid sequence, at least one polar aprotic solvent in an amount effective to denature double-stranded nucleotide sequences, an accelerating agent, and at least one additional component selected from the group consisting of: buffering agents, salts, chelating agents, detergents, and blocking agents, wherein less than 10% of the hybridization composition is formamide, wherein the polar aprotic solvent is not dimethyl sulfoxide (DMSO) and has lactone, sulfone, sulfite, and/or carbonate functional groups, wherein the at least one nucleic acid sequence is a FISH or CISH probe; wherein the at least one polar aprotic solvent comprises a polar aprotic solvent selected from the group consisting of:

wherein the accelerating agent is a polymer; and wherein said hybridization composition is configured to facilitate hybridization of said FISH or CISH probe component of the hybridization composition to a target nucleic acid in less than 8 hours.

2. The hybridization composition according to claim 1 , wherein the accelerating agent is present at a concentration from 1%-80%.

3. The hybridization composition according to claim 1 , wherein the accelerating agent is selected from the group consisting of PVP, heparin, dextran sulfate, and combinations thereof.

4. The hybridization composition according to claim 1 , wherein the accelerating agent is dextran sulfate.

5. The hybridization composition according to claim 4 , wherein the composition comprising up to 10% dextran sulfate.

6. The hybridization composition according to claim 4 , wherein the composition comprises up to 20% dextran sulfate.

7. The hybridization composition according to claim 1 , wherein the concentration of polar aprotic solvent is 5% to 10% (v/v).

8. The hybridization composition according to claim 1 , wherein the concentration of polar aprotic solvent is 10% to 20% (v/v).

9. The hybridization composition according to claim 1 , wherein the concentration of polar aprotic solvent is 20% to 30% (v/v).

10. The hybridization composition according to claim 1 , with the proviso that the composition does not contain formamide.

11. The hybridization composition according to claim 1 , wherein the composition contains less than 2% formamide.

12. The hybridization composition according to claim 1 wherein the composition contains less than 1% formamide.

13. The hybridization composition according to claim 1 , wherein the polar aprotic solvent is selected from the group consisting of: 2,3-butylene carbonate, γ-butyrolactone, caprolactone (epsilon), chloro maleic anhydride, chloroethylene carbonate, chloronitromethane, citraconic anhydride, crotonlactone, 5-cyano-2-thiouracil, dimethyl sulfate, dimethyl sulfone, diphenyl sulfone, ethanesulfonylchloride, ethylene carbonate, ethylene glycol sulfate, glycol sulfite, methyl alpha bromo tetronate, methyl phenyl sulfone, methyl sulfolane, methyl-4-toluenesulfonate, N-phenyl sydnone, phthalic anhydride, 1,3-propane sultone, β-propiolactone, propylene carbonate, saccharin, sulfanilamide, sulfolane, trimethylene sulfide-dioxide, and trimethylene sulfite.

14. The hybridization composition according to claim 1 , wherein the hybridization composition comprises at least one buffering agent and at least one salt.

15. The hybridization composition according to claim 14 , wherein the salt is NaCl and/or the buffering agent is phosphate buffer.

16. The hybridization composition according to claim 15 , wherein the accelerating agent is dextran sulfate present at a concentration of 10% to 40%, and wherein the NaCl is present at a concentration up to 1200 mM, and/or the phosphate buffer is present at a concentration up to 50 mM.

17. The hybridization composition according to claim 15 , wherein the dextran sulfate is present at a concentration of 10% to 30%, the NaCl is present at a concentration of 300 mM to 600 mM, and/or the phosphate buffer is present at a concentration of 5 mM to 20 mM.

18. The hybridization composition according to claim 1 , wherein the at least one nucleic acid sequence is a FISH DNA probe.

19. The hybridization composition according to claim 1 , wherein the at least one nucleic acid sequence comprises a detectable label.

Assignments (3)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 26, 2023
From: MATTHIESEN, STEEN HAUGE; PETERSEN, KENNETH H.; POULSEN, TIM SVENSTRUP; HANSEN, CHARLES M.
To: DAKO DENMARK A/S
Reel/Frame 065359/0410 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 26, 2023
From: DAKO DENMARK APS
To: AGILENT TECHNOLOGIES, INC.
Reel/Frame 065360/0336 →
CHANGE OF NAME Recorded Oct 26, 2023
From: DAKO DENMARK A/S
To: DAKO DENMARK APS
Reel/Frame 065379/0741 →
Priority Claims (2)
DK PA200800727 · May 27, 2008 · national
DK PA200900278 · Feb 27, 2009 · national
Continuity (5)
Continuation 17473524 · Sep 13, 2021
Continuation 12994492
Provisional Application 61155683 · Feb 26, 2009
Provisional Application 61056089 · May 27, 2008
Related Publication 20240124922A1 · Apr 18, 2024
References Cited (400)
US 4652517A · Scholl et al. · 1987 [cited by applicant]
US 4886741A · Schwartz · 1989 [cited by applicant]
US 4888278A · Singer et al. · 1989 [cited by applicant]
US 4996359A · Kesling, Jr. et al. · 1991 [cited by applicant]
US 5106730A · Van Ness et al. · 1992 [cited by applicant]
US 5132207A · Kohne et al. · 1992 [cited by applicant]
US 5382285A · Morrison · 1995 [cited by applicant]
US 5432065A · Fuller · 1995 [cited by applicant]
US 5521061A · Bresser et al. · 1996 [cited by applicant]
US 5525492A · Hill · 1996 [cited by applicant]
US 5527675A · Coull et al. · 1996 [cited by applicant]
US 5539082A · Nielsen et al. · 1996 [cited by applicant]
US 5582985A · Thompson · 1996 [cited by applicant]
US 5623049A · Obberding et al. · 1997 [cited by applicant]
US 5633129A · Karger et al. · 1997 [cited by applicant]
US 5650148A · Gage et al. · 1997 [cited by applicant]
US 5705333A · Shah et al. · 1998 [cited by applicant]
US 5714331A · Buchardt et al. · 1998 [cited by applicant]
US 5718915A · Virtanen et al. · 1998 [cited by applicant]
US 5736336A · Buchardt et al. · 1998 [cited by applicant]
US 5750340A · Kim · 1998 [cited by examiner]
US 5766855A · Buchardt et al. · 1998 [cited by applicant]
US 5773571A · Nielsen et al. · 1998 [cited by applicant]
US 5786461A · Buchardt et al. · 1998 [cited by applicant]
US 5837459A · Berg et al. · 1998 [cited by applicant]
US 5856089A · Wang et al. · 1999 [cited by applicant]
US 5858671A · Jones · 1999 [cited by applicant]
US 5869237A · Ward et al. · 1999 [cited by applicant]
US 5891625A · Buchardt et al. · 1999 [cited by applicant]
US 5919894A · Schubart · 1999 [cited by applicant]
US 5925744A · Haner et al. · 1999 [cited by applicant]
US 5962227A · Hedrick et al. · 1999 [cited by applicant]
US 5972610A · Buchardt et al. · 1999 [cited by applicant]
US 5986053A · Ecker et al. · 1999 [cited by applicant]
US 6107470A · Nielsen et al. · 2000 [cited by applicant]
US 6201103B1 · Nielsen et al. · 2001 [cited by applicant]
US 6203977B1 · Ward et al. · 2001 [cited by applicant]
US 6228982B1 · Norden et al. · 2001 [cited by applicant]
US 6331618B1 · Bloch et al. · 2001 [cited by applicant]
US 6344315B1 · Gray et al. · 2002 [cited by applicant]
US 6357163B1 · Buchardt et al. · 2002 [cited by applicant]
US 6361940B1 · Van Ness et al. · 2002 [cited by applicant]
US 6475720B1 · Gray et al. · 2002 [cited by applicant]
US 6555670B1 · Aizawa et al. · 2003 [cited by applicant]
US 6586176B1 · Trnovsky et al. · 2003 [cited by applicant]
US 6656685B2 · Utermohlen et al. · 2003 [cited by applicant]
US 6656734B1 · Bischoff · 2003 [cited by examiner]
US 6670461B1 · Wengel et al. · 2003 [cited by applicant]
US 6716625B1 · Selitrennikoff et al. · 2004 [cited by applicant]
US 6794499B2 · Wengel et al. · 2004 [cited by applicant]
US 6949368B2 · Chakrabarti et al. · 2005 [cited by applicant]
US 7034133B2 · Wengel et al. · 2006 [cited by applicant]
US 7105294B2 · Van Dongen et al. · 2006 [cited by applicant]
US 7329491B2 · Kirchgesser et al. · 2008 [cited by applicant]
US 7374945B2 · Becker · 2008 [cited by applicant]
US 7396508B1 · Richards et al. · 2008 [cited by applicant]
US 7572582B2 · Wengel et al. · 2009 [cited by applicant]
US 7655422B2 · Adler et al. · 2010 [cited by applicant]
US 7750208B2 · Huang et al. · 2010 [cited by applicant]
US 7867443B2 · Key et al. · 2011 [cited by applicant]
US 7901634B2 · Testa et al. · 2011 [cited by applicant]
US 8034909B2 · Wengel et al. · 2011 [cited by applicant]
US 8080644B2 · Wengel et al. · 2011 [cited by applicant]
US 8153365B2 · Wengel et al. · 2012 [cited by applicant]
US 8211385B2 · Testa et al. · 2012 [cited by applicant]
US 8632739B2 · Testa et al. · 2014 [cited by applicant]
US 8877144B2 · Poulsen et al. · 2014 [cited by applicant]
US 9133507B2 · Testa et al. · 2015 [cited by applicant]
US 9182323B2 · Poulsen et al. · 2015 [cited by applicant]
US 9297035B2 · Matthiesen et al. · 2016 [cited by applicant]
US 9303287B2 · Matthiesen · 2016 [cited by applicant]
US 9309562B2 · Matthiesen · 2016 [cited by applicant]
US 9388456B2 · Matthiesen · 2016 [cited by applicant]
US 9388466B2 · Chen et al. · 2016 [cited by applicant]
US 10031052B2 · Poulsen et al. · 2018 [cited by applicant]
US 10202638B2 · Matthiesen · 2019 [cited by applicant]
US 10295442B2 · Matthiesen et al. · 2019 [cited by applicant]
US 10416051B2 · Poulsen et al. · 2019 [cited by applicant]
US 10662465B2 · Matthiesen · 2020 [cited by applicant]
US 10996145B2 · Poulsen et al. · 2021 [cited by applicant]
US 11118214B2 · Matthiesen et al. · 2021 [cited by applicant]
US 11118226B2 · Matthiesen · 2021 [cited by applicant]
US 20010000487A1 · Essenfeld et al. · 2001 [cited by applicant]
US 20010007748A1 · An et al. · 2001 [cited by applicant]
US 20010009766A1 · Bard et al. · 2001 [cited by applicant]
US 20010010936A1 · Richards et al. · 2001 [cited by applicant]
US 20010011131A1 · Luyten et al. · 2001 [cited by applicant]
US 20010018512A1 · Blanchard · 2001 [cited by applicant]
US 20010026919A1 · Chenchik et al. · 2001 [cited by applicant]
US 20010027567A1 · Federoff · 2001 [cited by applicant]
US 20010056203A1 · Sezi et al. · 2001 [cited by applicant]
US 20020006652A1 · Danielsen et al. · 2002 [cited by applicant]
US 20020019001A1 · Light · 2002 [cited by applicant]
US 20020058278A1 · Stefano et al. · 2002 [cited by applicant]
US 20020065224A1 · Bender et al. · 2002 [cited by applicant]
US 20020076809A1 · Steinmeyer et al. · 2002 [cited by applicant]
US 20020102554A1 · Utermohlen et al. · 2002 [cited by applicant]
US 20020119465A1 · Zhao et al. · 2002 [cited by applicant]
US 20020127569A1 · Weisburg et al. · 2002 [cited by applicant]
US 20020137035A1 · Stender et al. · 2002 [cited by applicant]
US 20020150904A1 · Bi et al. · 2002 [cited by applicant]
US 20020164589A1 · Taylor · 2002 [cited by applicant]
US 20020164614A1 · Becker · 2002 [cited by applicant]
US 20020182613A1 · Mirkin et al. · 2002 [cited by applicant]
US 20020197629A1 · Gjerde et al. · 2002 [cited by applicant]
US 20020198366A1 · Ashkenazi et al. · 2002 [cited by applicant]
US 20030039992A1 · Chakrabarti et al. · 2003 [cited by applicant]
US 20030087237A1 · Hong et al. · 2003 [cited by applicant]
US 20030108903A1 · Wang et al. · 2003 [cited by applicant]
US 20030125312A1 · Rocha et al. · 2003 [cited by applicant]
US 20030134279A1 · Isola et al. · 2003 [cited by applicant]
US 20030175852A1 · Kalra et al. · 2003 [cited by applicant]
US 20030203391A1 · Sana et al. · 2003 [cited by applicant]
US 20040029184A1 · Gourevitch · 2004 [cited by applicant]
US 20040030093A1 · Sakurai et al. · 2004 [cited by applicant]
US 20040048376A1 · Chabot et al. · 2004 [cited by applicant]
US 20040053222A1 · Storhoff et al. · 2004 [cited by applicant]
US 20040096856A1 · Garimella et al. · 2004 [cited by applicant]
US 20040096967A1 · Gryseels et al. · 2004 [cited by applicant]
US 20040101946A1 · Hovanec · 2004 [cited by applicant]
US 20040106109A1 · Belly et al. · 2004 [cited by applicant]
US 20040122101A1 · Miller et al. · 2004 [cited by applicant]
US 20040157218A1 · Collis et al. · 2004 [cited by applicant]
US 20040210967A1 · Chen et al. · 2004 [cited by applicant]
US 20040219546A1 · Sakaki et al. · 2004 [cited by applicant]
US 20040224343A1 · Han et al. · 2004 [cited by applicant]
US 20040241666A1 · Amorese et al. · 2004 [cited by applicant]
US 20040248790A1 · Hinuma et al. · 2004 [cited by applicant]
US 20040268439A1 · Cheng et al. · 2004 [cited by applicant]
US 20050014154A1 · Weizenegger · 2005 [cited by applicant]
US 20050042627A1 · Chakrabarti et al. · 2005 [cited by applicant]
US 20050064472A1 · Shekar et al. · 2005 [cited by applicant]
US 20050079535A1 · Kirchgesser et al. · 2005 [cited by applicant]
US 20050191657A1 · Demorest et al. · 2005 [cited by applicant]
US 20050234236A1 · Kertesz et al. · 2005 [cited by applicant]
US 20050250094A1 · Storhoff et al. · 2005 [cited by applicant]
US 20050266459A1 · Poulsen et al. · 2005 [cited by applicant]
US 20050287548A1 · Bao et al. · 2005 [cited by applicant]
US 20060024751A1 · May et al. · 2006 [cited by applicant]
US 20060030541A1 · Garcia et al. · 2006 [cited by applicant]
US 20060040293A1 · Salonen et al. · 2006 [cited by applicant]
US 20060147957A1 · Qian et al. · 2006 [cited by applicant]
US 20060148846A1 · Orwat et al. · 2006 [cited by applicant]
US 20060160069A1 · Chau et al. · 2006 [cited by applicant]
US 20060191657A1 · Spence et al. · 2006 [cited by applicant]
US 20060270595A1 · Jullien et al. · 2006 [cited by applicant]
US 20070141583A1 · Li et al. · 2007 [cited by applicant]
US 20070148657A1 · Myerson et al. · 2007 [cited by applicant]
US 20070166641A1 · Shimizu et al. · 2007 [cited by applicant]
US 20070207482A1 · Church et al. · 2007 [cited by applicant]
US 20070220629A1 · Walsh et al. · 2007 [cited by applicant]
US 20070243545A1 · Kilpatrick et al. · 2007 [cited by applicant]
US 20080044385A1 · Nishi et al. · 2008 [cited by applicant]
US 20080050393A1 · Tang et al. · 2008 [cited by applicant]
US 20080076923A1 · Belogi et al. · 2008 [cited by applicant]
US 20080096262A1 · Kobayashi et al. · 2008 [cited by applicant]
US 20080108810A1 · Kertesz et al. · 2008 [cited by applicant]
US 20080188575A1 · Gaspar Martinho et al. · 2008 [cited by applicant]
US 20080220451A1 · Adler et al. · 2008 [cited by applicant]
US 20080227653A1 · Fodor et al. · 2008 [cited by applicant]
US 20080234474A1 · Braman et al. · 2008 [cited by applicant]
US 20080269502A1 · Gantz et al. · 2008 [cited by applicant]
US 20080318226A1 · Usui et al. · 2008 [cited by applicant]
US 20090123913A1 · Barany et al. · 2009 [cited by applicant]
US 20090186357A1 · Mauk et al. · 2009 [cited by applicant]
US 20090197346A1 · Winkler et al. · 2009 [cited by applicant]
US 20090221429A1 · Fujimoto et al. · 2009 [cited by applicant]
US 20090294305A1 · Bekki et al. · 2009 [cited by applicant]
US 20100047671A1 · Chiang et al. · 2010 [cited by applicant]
US 20100136542A1 · Lee et al. · 2010 [cited by applicant]
US 20100196902A1 · Pestano et al. · 2010 [cited by applicant]
US 20100243451A1 · Latham et al. · 2010 [cited by applicant]
US 20100279283A1 · Raghunath et al. · 2010 [cited by applicant]
US 20110229975A1 · Matthiesen et al. · 2011 [cited by applicant]
US 20110250698A1 · Pollner et al. · 2011 [cited by applicant]
US 20110262930A1 · Deleersnijder et al. · 2011 [cited by applicant]
US 20110281263A1 · Matthiesen et al. · 2011 [cited by applicant]
US 20110287951A1 · Emmert-Buck et al. · 2011 [cited by applicant]
US 20120309951A1 · Utermohlen et al. · 2012 [cited by applicant]
US 20120331587A1 · Lassen et al. · 2012 [cited by applicant]
US 20130017176A1 · Hosoda et al. · 2013 [cited by applicant]
US 20130040294A1 · Matthiesen · 2013 [cited by applicant]
US 20130072535A1 · Stierli et al. · 2013 [cited by applicant]
US 20130156695A1 · Sprecher et al. · 2013 [cited by applicant]
US 20130230918A1 · Wakamiya · 2013 [cited by applicant]
US 20140017704A1 · Casta et al. · 2014 [cited by applicant]
US 20140120535A1 · Aurich-Costa et al. · 2014 [cited by applicant]
US 20140242589A1 · Matthiesen · 2014 [cited by applicant]
US 20140256585A1 · McCoy · 2014 [cited by applicant]
US 20170283805A1 · Bonci et al. · 2017 [cited by applicant]
US 20190249235A1 · Matthiesen · 2019 [cited by applicant]
US 20200232018A1 · Matthiesen · 2020 [cited by applicant]
US 20200299769A1 · Matthiesen · 2020 [cited by applicant]
US 20220064716A1 · Hansen et al. · 2022 [cited by applicant]
US 20220154263A1 · Matthiesen · 2022 [cited by applicant]
US 20220162683A1 · Matthiesen · 2022 [cited by applicant]
CN 101124337A · 2008 [cited by applicant]
CN 103966321A · 2014 [cited by applicant]
CN 104032033A · 2014 [cited by applicant]
EA 005581B1 · 2005 [cited by applicant]
EP 0261955A2 · 1988 [cited by applicant]
EP 1013772A1 · 2000 [cited by applicant]
WO 9102088A1 · 1991 [cited by applicant]
WO 9204341A1 · 1992 [cited by applicant]
WO 9207956A1 · 1992 [cited by applicant]
WO 9207959A1 · 1992 [cited by applicant]
WO 9402638A1 · 1994 [cited by applicant]
WO 9402639A1 · 1994 [cited by applicant]
WO 9604000A1 · 1996 [cited by applicant]
WO 9914226A2 · 1999 [cited by applicant]
WO 9927103A1 · 1999 [cited by applicant]
WO 0006773A1 · 2000 [cited by applicant]
WO 0069899A1 · 2000 [cited by applicant]
WO 0166804A2 · 2001 [cited by applicant]
WO 02061137A2 · 2002 [cited by applicant]
WO 02088396A2 · 2002 [cited by applicant]
WO 03014398A2 · 2003 [cited by applicant]
WO 03027328A2 · 2003 [cited by applicant]
WO 03054209A2 · 2003 [cited by applicant]
WO 2006007841A2 · 2006 [cited by applicant]
WO 2006023919A2 · 2006 [cited by applicant]
WO 2006066039A2 · 2006 [cited by applicant]
WO 2006093150A1 · 2006 [cited by applicant]
WO 2006117596A2 · 2006 [cited by applicant]
WO 2007019432A2 · 2007 [cited by applicant]
WO 2007019492A2 · 2007 [cited by applicant]
WO 2007037314A1 · 2007 [cited by applicant]
WO 2007037341A1 · 2007 [cited by applicant]
WO 2007058326A1 · 2007 [cited by applicant]
WO 2007109441A2 · 2007 [cited by applicant]
WO 2007109941A1 · 2007 [cited by applicant]
WO 2009074154A2 · 2009 [cited by applicant]
WO 2009144561A2 · 2009 [cited by applicant]
WO 2009144581A1 · 2009 [cited by applicant]
WO 2009147537A2 · 2009 [cited by applicant]
WO 2010097655A1 · 2010 [cited by applicant]
WO 2010097656A1 · 2010 [cited by applicant]
WO 2010097707A1 · 2010 [cited by applicant]
WO 2011067678A2 · 2011 [cited by applicant]
WO 2011153354A1 · 2011 [cited by applicant]
WO 2013046033A1 · 2013 [cited by applicant]
WO 2014131906A1 · 2014 [cited by applicant]
WO 2014176575A1 · 2014 [cited by applicant]
WO 2015103287A2 · 2015 [cited by applicant]
WO 2015184144A1 · 2015 [cited by applicant]
WO 2016125091A1 · 2016 [cited by applicant]
The European Agency for the Eval, et al., “2-Pyrrolidone,” Summary Report, Committee for Veterinary Medical Products, Retrieved from the Internet: <URL: www.ema.europa.eu/docs/en_GB/document_library/Maximum_Residue_Limi… [cited by applicant]
Tkachuk, D C. et al., “Detection of BCR-ABL Fusion In Chronic Myelogeneous Leukemia By In Situ Hybridization,” Science, vol. 250, Issue 4980, Oct. 26, 1990, 559-562. [cited by applicant]
Toyozo, Takahashi et al., “Principal Assay Method in Infection Diagnosis, Chapter 3. Diagnosis of Infection by DNA Probe,” Applied DNA Probe Technology, Feb. 5, 1988, 8 pages. [cited by applicant]
Tsuruoka, Makoto et al., “Rapid Hybridization at High Salt Concentration and Detection of Bacterial DNA Using Fluorescence Polarization,” Combinatorial Chemistry & High Throughput Screening, vol. 6, No. 3, 2003, 225-234. [cited by applicant]
USPTO, “Notice of Allowance mailed on Apr. 22, 2022,” U.S. Appl. No. 16/240,448, 24 pages. [cited by applicant]
Wahl, Geoffrey M. et al., “Efficient Transfer of Large DNA Fragments From Agarose Gels to Diazobenzyloxymethyl—Paper and Rapid Hybridization by Using Dextran Sulfate,” Proc. Natl. Acad. Sci. USA. Biochemistry, vol. 76, … [cited by applicant]
Wang, Pei et al., “Simultaneous Detection and Differentiation of [cited by applicant]
Webpage, et al., “OPTIM Synthetic Glycerine—Vapor Pressure and Boiling Point,” Webpage, Jun. 28, 2007, 1 page. [cited by applicant]
Wikipedia, et al., “2-Pyrrolidone,” Webpage, Nov. 1, 2012, 3 pages. [cited by applicant]
Wikipedia, et al., “Polyalkylenglycolether,” noch ältere version von Nov. 12, 2006, Nov. 12, 2006, 2 pages. [cited by applicant]
Wikipedia, et al., “Polyalkylenglycolether,” ältere Version von Feb. 18, 2015, Feb. 18, 2015, 4 pages. [cited by applicant]
Wikipedia, et al., “Polyalkylenglycolether,” Wikipedia, May 25, 2021, 4 pages. [cited by applicant]
Wikipedia, et al., “Polysorbate 20,” Wikipedia, 3 pages. [cited by applicant]
Wikipedia, et al., “Solvent,” Wikipedia, Apr. 17, 2019, 15 pages. [cited by applicant]
Wikipedia, et al., “Triton X-100,” Wikipedia, Mar. 12, 2021, 4 pages. [cited by applicant]
Wilkinson, David G. et al., “Detection of Messenger RNA by in Situ Hybridization to Tissue Sections and Whole Mounts,” Methods in Enzymology, vol. 225, No. 361, 1993, 361-373. [cited by applicant]
Winship, Peter R. et al., “An Improved Method For Directly Sequencing PCR Amplified Material Using Dimethyl Sulphoxide,” Nucleic Acids Research, vol. 17, No. 3, 1989, 1266 pages. [cited by applicant]
Woenckhaus, Matthias et al., “Multitarget FISH And LOH Analyses at Chromosome 3p in Non-Small Cell Lung Cancer and Adjacent Bronchial Epithelium,” Am. J. of Clin. Pathol., vol. 123, No. 5, 2005, 752-761. [cited by applicant]
Xi, Chuanwu et al., “Use of DNA and Peptide Nucleic Acid Molecular Beacons for Detection and Quantification of rRNA in Solution and in Whole Cells,” Applied and Environmental Microbiology, vol. 69, No. 9, Sep. 9, 2003, … [cited by applicant]
Yuan, Liudi et al., “7 Duxiu Knowledge, Dimethylsulfoxide and Hybridization and Reduce Tm,” Advanced Biochemistry and Molecular Biology Experiment Course, Mar. 31, 2006, 58-60. [cited by applicant]
Zhao, Jia et al., “Identification of BAC Clones for Array CGH in Human Cancer by FISH Technique,” Journal of Jilin University (Medical), vol. 34, No. 4, Jul. 28, 2006, 629-631. [cited by applicant]
EPO, “Communication Re: Patentee Response to Opposition mailed on Jan. 11, 2021,” Application No. 15184389.3, 24 pages. [cited by applicant]
EPO, “Communication to Opponent Concerning Maintenance mailed on Jul. 24, 2019,” Application No. 13164094.8, 4 pages. [cited by applicant]
EPO, “Communication to Opponent Concerning Maintenance mailed on Oct. 28, 2019,” Application No. 09754209.6, 2 pages. [cited by applicant]
EPO, “Decision Revoking the European Patent mailed on Nov. 29, 2019,” Application No. 12775027.1, 46 pages. [cited by applicant]
EPO, “Decision to maintain the European Patent (Opponent) mailed on Oct. 24, 2019,” Application No. 13164094.8, 1 page. [cited by applicant]
EPO, “Decision to maintain the European patent in amended form (Art. 101(3)(a) EPC) mailed on Dec. 15, 2022,” Application No. 15184389.3, 1 page. [cited by applicant]
EPO, “Decision to maintain the European patent in amended form mailed on Jan. 23, 2020,” Application No. 09754209.6, 1 page. [cited by applicant]
EPO, “Decision to maintain the European patent in amended form mailed on Jan. 23, 2020 (Opponent),” Application No. 09754209.6, 1 page. [cited by applicant]
EPO, “Decision to maintain the European Patent mailed on Oct. 24, 2019,” Application No. 13164094.8, 1 page. [cited by applicant]
EPO, “Declaration of Dr: Jens Mollerup,” Curriculum Vitae of Dr. Jens Mollerup & Histology FISH Accessory Kit, Code K5799 instructions for use, 43 pages. [cited by applicant]
EPO, “Electronic Versions of the Handwritten Amendments (Written Submission) submitted on Dec. 19, 2018,” Application No. 13164094.8, 26 pages. [cited by applicant]
EPO, “Electronic Versions of the Handwritten Amendments submitted on May 31, 2019,” Application No. 09754209.6, 14 pages. [cited by applicant]
EPO, “English Translation of Auxiliary Request 1 (Tracked & Clean) filed by Patentee on Dec. 18, 2020,” Application No. 15184389.3, 4 pages. [cited by applicant]
EPO, “English Translation of Auxiliary Request 2 (Tracked & Clean) filed by Patentee on Dec. 18, 2020,” Application No. 15184389.3, 4 pages. [cited by applicant]
EPO, “English Translation of Auxiliary Request 3 (Tracked & Clean) filed by Patentee on Dec. 18, 2020,” Application No. 15184389.3, 4 pages. [cited by applicant]
EPO, “English Translation of Auxiliary Request 4 (Tracked & Clean) filed by Patentee on Dec. 18, 2020,” Application No. 15184389.3, 4 pages. [cited by applicant]
EPO, “English Translation of Auxiliary Request 5 (Tracked & Clean) filed by Patentee on Dec. 18, 2020,” Application No. 15184389.3, 4 pages. [cited by applicant]
EPO, “English Translation of Main Request (Tracked and Clean) filed by Patentee on Dec. 18, 2020,” Application No. 15184389.3, 4 pages. [cited by applicant]
EPO, “English Translation of Summons to Oral Proceedings pursuant to Rule 115(1) EPC mailed on Apr. 8, 2021,” Application No. 15184389.3, 11 pages. [cited by applicant]
EPO, “EPO Comm Re Refuse to File the Request to Correct the Minutes of Oral Proceedings mailed on Jul. 4, 2022,” Application No. 15184389.3, 2 pages. [cited by applicant]
EPO, “EPO Communication Re: Letter from Proprietor dated Jan. 7, 2022 mailed on Jan. 12, 2022,” Application No. 15184389.3, 93 pages. [cited by applicant]
EPO, “EPO Notification Re Extension of Response Deadline Filed by Patentee mailed on Nov. 5, 2020,” Application No. 15184389.3, 4 pages. [cited by applicant]
EPO, “EPO Refund of Appeal Fee mailed on Aug. 5, 2022,” Application No. 15184389.3, 1 page. [cited by applicant]
EPO, “EPO Request for Documents Pursuant Rule 83 EPC/ Request for Translation Pursuant Rule 3(3) EPC to Opponent mailed on Oct. 20, 2017,” Application No. 09754209.6, 1 page. [cited by applicant]
EPO, “EPO Submission from the Patentee Filed on Mar. 7, 2022,” Application No. 15184389.3, 21 pages. [cited by applicant]
EPO, “Extended European Search Report mailed on Aug. 14, 2013,” Application No. 13164099.7, 5 pages. [cited by applicant]
EPO, “Extended European Search Report mailed on Aug. 19, 2020,” Application No. 20167307.6, 7 pages. [cited by applicant]
EPO, “Extended European Search Report mailed on Aug. 8, 2013,” Application No. 13164094.8, 5 pages. [cited by applicant]
EPO, “Extended European Search Report mailed on Jun. 6, 2018,” Application No. 18165184.5, 8 pages. [cited by applicant]
EPO, “File Request to Correct the Minutes of Oral Proceedings filed on Jun. 23, 2022,” Application No. 15184389.3, 4 pages. [cited by applicant]
EPO, “File Request to Withdraw the Appeal Filed on Jul. 27, 2022,” Application 15184389.3, 3 pages. [cited by applicant]
EPO, “Further Response to Patentee's Submission (dated Jan. 7, 2022) Filed on Mar. 1, 2022,” Application No. 15184389.3, 8 pages. [cited by applicant]
EPO, “Information About the Result of Oral Proceedings mailed on May 7, 2019,” Application No. 09754209.6, 10 pages. [cited by applicant]
EPO, “Information About the Result of Oral Proceedings mailed on Nov. 4, 2019,” Application No. 12775027.1, 9 pages. [cited by applicant]
EPO, “Information About the Result of Oral Proceedings mailed on Nov. 30, 2018,” Application No. 13164094.8, 6 pages. [cited by applicant]
EPO, “Interlocutory Decision in Opposition Proceedings (Art. 101(3)(a) and 106(2) EPC) mailed on Apr. 14, 2022,” Application No. 15184389.3, 60 pages. [cited by applicant]
EPO, “Interlocutory Decision in Opposition Proceedings (Art. 101(3)(a) and 106(2) EPC) mailed on Jul. 1, 2019,” Application No. 09754209.6, 66 pages. [cited by applicant]
EPO, “Interlocutory Decision in Opposition Proceedings (Art. 101(3)(a) and 106(2) EPC) mailed on Mar. 5, 2019,” Application No. 13164094.8, 40 pages. [cited by applicant]
EPO, “Internal Form—Opposition/Addressees mailed on Jul. 1, 2019,” Application No. 09754209.6, 2 pages. [cited by applicant]
EPO, “Internal Form—Opposition/Addressees mailed on Mar. 5, 2019,” Application No. 13164094.8, 2 pages. [cited by applicant]
EPO, “Main Request (Tracked and Clean) filed by Patentee on Dec. 18, 2020,” Application No. 15184389.3, 8 pages. [cited by applicant]
EPO, “Maintenance Of The Patent With The Documents Specified In The Final Decision mailed on Jul. 17, 2019,” Application No. 13164094.8, 1 page. [cited by applicant]
EPO, “Maintenance of the Patent with the Documents Specified in the Final Decision mailed on Oct. 17, 2019,” Application No. 09754209.6, 1 page. [cited by applicant]
EPO, “Minutes of the Oral Proceedings (Opposition Division)—Conclusion of the Proceedings mailed on Mar. 5, 2019,” Application No. 13164094.8, 3 pages. [cited by applicant]
EPO, “Notice of Appeal Filed on Jun. 24, 2022,” Application No. 15184389.3, 5 pages. [cited by applicant]
EPO, “Notice of Opposition by Opponent filed on Apr. 18, 2018,” Application No. 12775027.1, 34 pages. [cited by applicant]
EPO, “Notice of Opposition by Opponent filed on Apr. 20, 2017,” Application No. 13164094.8, 37 pages. [cited by applicant]
EPO, “Notice of Opposition by Opponent filed on Oct. 11, 2017,” Application No. 09754209.6, 31 pages. [cited by applicant]
EPO, “Oppodrex with Handwritten Amendments mailed on Jul. 1, 2019,” Application No. 09754209.6, 39 pages. [cited by applicant]
EPO, “Oppodrex with Handwritten Amendments mailed on Mar. 5, 2019,” Application No. 13164094.8, 50 pages. [cited by applicant]
EPO, “Advice of Delivery mailed on May 29, 2018,” Application No. 13164094.8, 1 page. [cited by applicant]
EPO, “Advice of Delivery mailed on May 30, 2018,” Application No. 13164094.8, 2 pages. [cited by applicant]
EPO, “Annex 1—Claim Synopsis,” Opposition to European Patent No. 3133166, 2 pages. [cited by applicant]
EPO, “Auxiliary Request 1 (Tracked & Clean) filed by Patentee on Dec. 18, 2020,” Application No. 15184389.3, 8 pages. [cited by applicant]
EPO, “Auxiliary Request 1-6 (Marked & Clean) filed by Patentee on Mar. 7, 2022,” Application No. 15184389.3, 59 pages. [cited by applicant]
EPO, “Auxiliary Request 2 (Tracked & Clean) filed by Patentee on Dec. 18, 2020,” Application No. 15184389.3, 8 pages. [cited by applicant]
EPO, “Auxiliary Request 3 (Tracked & Clean) filed by Patentee on Dec. 18, 2020,” Application No. 15184389.3, 8 pages. [cited by applicant]
EPO, “Auxiliary Request 4 (Tracked & Clean) filed by Patentee on Dec. 18, 2020,” Application No. 15184389.3, 8 pages. [cited by applicant]
EPO, “Auxiliary Request 5 (Tracked & Clean) filed by Patentee on Dec. 18, 2020,” Application No. 15184389.3, 8 pages. [cited by applicant]
EPO, “Auxiliary Request 7-10 (Marked & Clean) filed by Patentee on Mar. 7, 2022,” Application No. 15184389.3, 33 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Feb. 5, 2018,” Application No. 09754209.6, 4 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Feb. 7, 2018,” Application No. 13164094.8, 14 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Jan. 7, 2019,” Application No. 13164094.8, 27 pages. [cited by applicant]
EPO, “Brief communication—Opposition Proceedings mailed on Jan. 8, 2018,” Application No. 09754209.6, 3 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Jul. 1, 2019,” Application No. 09754209.6, 15 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Mar. 11, 2019,” Application No. 09754209.6, 2 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Mar. 11, 2019,” Application No. 09754209.6, 83 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Mar. 13, 2019,” Application No. 09754209.6, 23 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on May 25, 2018,” Application No. 13164094.8, 16 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on May 25, 2018,” Application No. 09754209.6, 64 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on May 3, 2019,” Application No. 09754209.6, 51 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Nov. 5, 2019,” Application No. 12775027.1, 1 page. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Nov. 13, 2018,” Application No. 13164094.8, 6 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Nov. 30, 2018,” Application No. 13164094.8, 124 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Oct. 1, 2019,” Application No. 09754209.6, 7 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Oct. 16, 2017,” Application No. 13164094.8, 67 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Oct. 2, 2018,” Application No. 12775027.1, 27 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Oct. 5, 2018,” Application No. 13164094.8, 77 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Oct. 5, 2018,” Application No. 13164094.8, 8 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Sep. 6, 2019,” Application No. 12775027.1, 2 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Sep. 9, 2019,” Application No. 12775027.1, 2 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Sep. 12, 2019,” Application No. 12775027.1, 6 pages. [cited by applicant]
EPO, “Brief Communication—Opposition Proceedings mailed on Sep. 16, 2019,” Application No. 12775027.1, 1 page. [cited by applicant]
EPO, “Brief Communication of Jan. 8, 2018 of the EPO in the Opposition Proceedings Including the Aforementioned Document D14 (Termed “D5” in the Examination and Opposition Proceedings of EP 2285979) mailed on Feb. 7, 20… [cited by applicant]
EPO, “Commencement of Proceedings Before the Board of Appeal mailed on Jul. 12, 2022,” Application No. 15184389.3, 2 pages. [cited by applicant]
EPO, “Communication of a Notice of Opposition—First Information to Patent Proprietor mailed on Apr. 24, 2018,” Application No. 12775027.1, 1 page. [cited by applicant]
EPO, “Communication of a Notice of Opposition—First Information to Patent Proprietor mailed on Apr. 26, 2017,” Application No. 13164094.8, 1 page. [cited by applicant]
EPO, “Communication of a Notice of Opposition—First Information to Patent Proprietor mailed on Oct. 20, 2017,” Application No. 09754209.6, 1 page. [cited by applicant]
EPO, “Communication of Bibliographic Data mailed on Oct. 28, 2019,” Application No. 09754209.6, 2 pages. [cited by applicant]
EPO, “Communication of Further Notices of Opposition Pursuant to Rule 79(2) EPC mailed on Jan. 15, 2018,” Application No. 09754209.6, 1 page. [cited by applicant]
EPO, “Communication of Further Notices of Opposition Pursuant to Rule 79(2) EPC mailed on May 30, 2017,” Application No. 13164094.8, 1 page. [cited by applicant]
EPO, “Communication of Further Notices of Opposition Pursuant to Rule 79(2) EPC mailed on May 8, 2018,” Application No. 12775027.1, 1 page. [cited by applicant]
EPO, “Communication of Notices of Opposition (R. 79(1) EPC) mailed on Jan. 15, 2018,” Application No. 09754209.6, 1 page. [cited by applicant]
EPO, “Communication of Notices of Opposition (R. 79(1) EPC) mailed on May 30, 2017,” Application No. 13164094.8, 1 page. [cited by applicant]
EPO, “Communication of Notices of Opposition (R. 79(1) EPC) mailed on May 8, 2018,” Application No. 12775027.1, 1 page. [cited by applicant]
EPO, “Communication Pursuant to Rule 164(2)(b) and Article 94(3) EPC mailed on Jun. 12, 2015,” Application No. 12775027.1, 6 pages. [cited by applicant]
EPO, “Communication Pursuant to Rule 82(2) EPC mailed on Sep. 14, 2022,” Application No. 15184389.3, 4 pages. [cited by applicant]
EPO, “Communication Pursuant to Rule 82(2) EPC to Pay Printing Fees and File Translations mailed on Jul. 24, 2019,” Application No. 13164094.8, 6 pages. [cited by applicant]
EPO, “Communication Pursuant to Rule 82(2) EPC to Pay Printing Fees and File Translations mailed on Oct. 28,” 2019, Application No. 09754209.6, 2 pages. [cited by applicant]
EPO, “Patentee Response to Opposition filed on Dec. 18, 2020 (Including English Translation),” Application No. 15184389.3, 45 pages. [cited by applicant]
EPO, “Patentee's Filings of the Adapted Description V1 dated Mar. 8, 2022,” Application No. 15184389.3, 7 pages. [cited by applicant]
EPO, “Patentee's Filings of the Adapted Description V2 dated Mar. 8, 2022,” Application No. 15184389.3, 8 pages. [cited by applicant]
EPO, “Patentee's Letter of Oct. 15, 2015 in the Examination Proceedings,” Application No. 12775027.1, 7 pages. [cited by applicant]
EPO, “Provision of the Minutes in accordance with Rule 124(4) EPC mailed on Apr. 14, 2022,” Application No. 15184389.3, 24 pages. [cited by applicant]
EPO, “Provision of the Minutes in Accordance with Rule 124(4) EPC mailed on Jul. 1, 2019,” Application No. 09754209.6, 21 pages. [cited by applicant]
EPO, “Provision of the Minutes in Accordance with Rule 124(4) EPC mailed on Mar. 5, 2019,” Application No. 13164094.8, 25 pages. [cited by applicant]
EPO, “Reply from the Opponent to Submission of Proprietor filed on Feb. 1, 2018,” Application No. 13164094.8, 13 pages. [cited by applicant]
EPO, “Reply of the Patent Proprietor to the Notice(s) of Opposition Filed on May 17, 2018,” Application No. 09754209.6, 67 pages. [cited by applicant]
EPO, “Reply of the Patent Proprietor to the Notice(s) of Opposition filed on Oct. 6, 2017,” Application No. 13164094.8, 66 pages. [cited by applicant]
EPO, “Reply of the Patent Proprietor to the Notice(s) of Opposition filed on Sep. 18, 2018,” Application No. 12775027.1, 26 pages. [cited by applicant]
EPO, “Request for Correction in Minutes of Oral Proceedings filed on Aug. 21, 2019,” Application No. 09754209.6, 3 pages. [cited by applicant]
EPO, “Request for Correction in Reference Number by Opponent filed on Jan. 23, 2018,” Application No. 09754209.6, 3 pages. [cited by applicant]
EPO, “Request for Documents Pursuant to Rule 83 EPC by Opponent filed on Jan. 2, 2018,” Application No. 09754209.6, 3 pages. [cited by applicant]
EPO, “Request for Interpreters by Opponent filed on Mar. 5, 2019,” Application No. 09754209.6, 1 page. [cited by applicant]
EPO, “Request for Interpreters by Opponent filed on Sep. 2, 2019,” Application No. 12775027.1, 3 pages. [cited by applicant]
EPO, “Request for Interpreters filed on Sep. 2, 2019,” Application No. 12775027.1, 1 page. [cited by applicant]
EPO, “Response to Brief Communication of Jan. 8, 2018 & Further to Notice of Opposition by Opponent filed on Jan. 26, 2018,” Application No. 09754209.6, 4 pages. [cited by applicant]
EPO, “Response to Communication Pursuant to Rule 82(2) EPC Filed on Oct. 11, 2019,” Application No. 13164094.8, 19 pages. [cited by applicant]
EPO, “Response to Patentee's Submission (dated Dec. 18, 2020) filed on Apr. 1, 2021,” Application No. 15184389.3, 19 pages. [cited by applicant]
EPO, “Response to Patentee's Submission (dated Jan. 7, 2022) Filed on Feb. 18, 2022,” Application No. 15184389.3, 13 pages. [cited by applicant]
EPO, “Response to Summons to Attend Oral Proceedings Filed on Dec. 20, 2021,” Application No. 15184389.3, 11 pages. [cited by applicant]
EPO, “Response to the Opponent's Letter of Feb. 1, 2018 filed on May 17, 2018,” Application No. 13164094.8, 15 pages. [cited by applicant]
EPO, “Statement by Dr Pierre Marggraf Rogalla,” Curriculum vitae: Dr. Pierre Marggraf-Rogalla (born Rogalla), 3 pages. [cited by applicant]
EPO, “Summons to Attend Oral Proceedings mailed on Aug. 21, 2018,” Application No. 09754209.6, 21 pages. [cited by applicant]
EPO, “Summons to Attend Oral Proceedings mailed on Feb. 11, 2019,” Application No. 12775027.1, 6 pages. [cited by applicant]
EPO, “Summons to Attend Oral Proceedings mailed on May 22, 2018,” Application No. 13164094.8, 37 pages. [cited by applicant]
EPO, “Summons to Attend Oral Proceedings Received by Opponent mailed on Aug. 21, 2018,” Application No. 09754209.6, 21 pages. [cited by applicant]
EPO, “Summons to Attend Oral Proceedings Received by Opponent mailed on Feb. 11, 2019,” Application No. 12775027.1, 6 pages. [cited by applicant]
EPO, “Summons to Attend Oral Proceedings Received by Opponent mailed on May 22, 2018,” Application No. 13164094.8, 37 pages. [cited by applicant]
EPO, “Summons to Oral Proceedings pursuant to Rule 115(1) EPC mailed on Apr. 8, 2021,” Application No. 15184389.3, 14 pages. [cited by applicant]
EPO, “Termination of the opposition proceedings with maintenance of patent mailed on Jan. 22, 2020,” Application No. 09754209.6, 1 page. [cited by applicant]
EPO, “Termination of the Opposition Proceedings with Maintenance of Patent mailed on Oct. 18, 2019,” Application No. 13164094.8, 1 page. [cited by applicant]
EPO, “Translation of the amended claims filed in French and German filed on Jan. 9, 2020,” Application No. 09754209.6, 12 pages. [cited by applicant]