DETAILED ACTION Notice of Pre-AIA or AIA Status The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA. Election/Restrictions Applicant’s election without traverse of the specie, a single affinity reagent (Claims1-5, 8-10, 13- 15, 17, 19, 22-24, 26 - 27, 34-36, 38, 40-44 and 51) in the reply filed on 10/31/2025 is acknowledged. Claims 6-7 and 48 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected species, drawn two or more affinity reagents, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 10/31/2025. Claims Status Claims 1-10, 13-15, 17, 19, 22-24, 26-27, 34-36, 38, 40-44, 48 and 51 are pending. Claims 6-7 and 48 are withdrawn. Claims 1-5, 8-10, 13-15, 17, 19, 22-24, 26-27, 34-36, 38, 40-44 and 51 are currently under examination . Priority This application is a 371 national phase application of PCT/US21/49944, filed on 09/10/2021, which claims benefit of U.S. Provisional Application Nos. 63/077,496, filed on 09/11/2020, and 63/196,953, filed on 06/ 04/2021. The priority date of claim s 1-5, 8-10, 13-15, 17, 19, 22-24, 26-27, 34-36, 38, 40-43 and 51 of claim set filed on 03/07/2023, is determined to be 09/11/2020. The priority date of claim 44 of claim set filed on 03/07/2023, is determined to be the filling date of PCT/US21/49944, filed on 09/10/2021. Specification The use of the terms CUTAC, CUT&Tag2for 1, CUT&Tag-direct and CUT&Tag (starting from Pg. 6-7), which are a trade names or a marks used in commerce, has been noted in this application. The term should be accompanied by the generic terminology; furthermore the term should be capitalized wherever it appears or, where appropriate, include a proper symbol indicating use in commerce such as ™, SM , or ® following the term . Although the use of trade names and marks used in commerce (i.e., trademarks, service marks, certification marks, and collective marks) are permissible in patent applications, the proprietary nature of the marks should be respected and every effort made to prevent their use in any manner which might adversely affect their validity as commercial marks. Claim Rejections - 35 USC § 112 The following is a quotation of 35 U.S.C. 112(b): (b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention. The following is a quotation of 35 U.S.C. 112 (pre-AIA), second paragraph: The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention. Claim s 13-14 and 26 rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention. Claim 13 is indefinite over the limitation “monovalent ionic concentrations”. It is unclear if the limitation is intended to claim monovalent, since Mg 2+ and Ca 2+ are divalent cation s , and generally used at working concentrations of 10mM or less, or whether the salt concentration is considered monovalent , where the salt may be NaCl or KCl . Claim 14 depends on claim 13 . Claim 13 is indefinite over the phrase “less than about" is vague and indefinite based on the unclear metes and bounds meant by the phrase. The phrase "less than” typically indicates a maximum point to not exceed. The phrase "less than", however, is controverted by the term "about" which implies that values above and below are permitted. Thus, the phrase "less than about" is indefinite where the metes and bounds of the term were not defined in the specification because it is not clear if “less than about” 10 mM would include concentration that were about 10 mM (e.g., 11 mM), because it is not clear if 11 mM would be less than 10 mM. Claim 26 is indefinite over the limitation “two different transposases”. As many transposases function as a dimer, it is unclear whether two different transposases is intended to characterize a dimer comprised of a molecule or complex formed by two identical units or if one of the two units is characteristically different ( e.g. comprising mutant(s) ) from the other unit. Claim Rejections - 35 USC § 102 The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action: A person shall be entitled to a patent unless – (a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention. Claim s are 1- 2, 4, 9, 13, 15, 17, 19, 22, 24, 26-27, 34-36, 38, 41-44, 48 and 51 are rejected under 35 U.S.C. 102 (a)(1) as being anticipated by Henikoff et al. (“ Henikoff ”; Patent App. Pub. WO 2019060907 A1, March 28, 2019). Interpretation: The limitation “optionally”, recited in claims 10, 15, 17, 23, 27, 36, 38, and 44, is interpreted as rendering the phrase thereafter as an optional limitation to the claim. Henikoff discloses “ A method for detecting the binding of a chromatin-associated factor of interest to a sequence of chromatin DNA in a cell, including: contacting a permeabilized cell or nucleus with a specific binding agent that specifically recognizes the chromatin-associated factor of interest, wherein the specific binding agent is linked to a nuclease that is inactive or an activatable transposome ; activating the nuclease or transposase, thereby excising the sequence of chromatin DNA bound to the chromatin- associated factor of interest; isolating the excised DNA; and determining the sequence of the excised DNA, thereby detecting binding of a chromatin-associated factor of interest to a sequence of chromatin DNA in the cell. .” (Abstract). Regarding claim 1, Henikoff teaches an in situ method for detecting a site of DNA accessibility in the chromatin of a cell, comprising: contacting a permeabilized cell with a first affinity reagent that specifically binds a nucleosome depleted region (NDR) marker, wherein the first affinity reagent is coupled to at least one transposome comprising: at least one transposase; and a transposon comprising: a first DNA molecule comprising a first transposase recognition site; and a second DNA molecule comprising a second transposase recognition site; activating the at least one transposase under low ionic conditions, thereby cleaving and tagging chromatin DNA with the first and second DNA molecules and excising a tagged DNA segment associated with the NDR marker; isolating the excised tagged DNA segment; and determining the nucleotide sequence of the excised tagged DNA segment, thereby detecting the site of DNA accessibility in the chromatin of the cell. (Para. 155) The teachings of Henikoff are documented above in the rejection of claim 1 under 35 U.S.C. 10 2 (a)(1) . Claims 2, 4, 9, 13, 15, 17, 19, 22, 24, 26-27, 34-36, 38, 41-44, 48 and 51 depend on claim 1. Claim 3 depends on claim 2, which depends on claim 1. Claim 3 depends on claim 2, which depends on claim 1. Claim 8 depends on claim 5. Claims 5-7 depend on claim 4, which depend on claim 1. Claim 10 depend on claim 9, which depends on claim 1. Claim 14 depends on claim 13, which depends on claim 1. Claim 23 depends on claim 22, which depends on claim 1. Claim 40 depends on claim 38, which depends on claim 1. Regarding claim 2-3, Henikoff teaches a method wherein the first affinity reagent is directly coupled to at least one transposase; and wherein the first affinity reagent and transposase are disposed in a fusion protein. (Para. 67) Regarding claim 4, Henikoff teaches a method wherein the first affinity reagent is indirectly coupled to the at least one transposase. (Para. 139) Regarding claim 5, Henikoff teaches a method wherein the transposase is linked to a specific binding agent that specifically binds the first affinity reagent. (Para. 139) Regarding claim 8, Henikoff teaches a method wherein the specific binding agent comprises protein A or protein G that specifically binds the first affinity reagent, the second affinity reagent and/or the third affinity reagent. (Para. 139) Regarding claim 9-10, Henikoff teaches a method wherein the first, second, and/or third affinity reagents independently is or comprises an antibody, an antibody- like molecule, a DARPin , an aptamer, a chromatin-binding protein, other specifically binding molecule, or a functional antigen-binding domain thereof; and wherein the antibody-like molecule is an antibody fragment and/or antibody derivative, optionally a single-chain antibody, a bispecific antibody, an Fab fragment, an F(ab)2fragment, a VHH fragment, a VNAR fragment, or a nanobody, optionally wherein the single-chain antibody is a single chain variable fragment ( scFv ), or a single-chain Fab fragment ( scFab ). (Para. 143; Para. 92) Regarding claim 13-14, Henikoff teaches a method wherein the low ionic conditions are characterized by monovalent ionic concentration of less than about 10 mM; and wherein the low ionic conditions are obtained by diluting liquid conditions of the transposase with a Mg ++ solution, removing liquid supernatant from the transposase and replacing it with a low ionic strength solution, and/or conducting a stringent (e.g., 300 mM) wash followed by adding a low ionic strength solution. ( Para. 223; Para. 887) Regarding claim 15, Henikoff teaches a method further comprising contacting the permeabilized cell with a polar compound prior to or during the step of activating the transposase under low ionic conditions . (Para. 223) Regarding claim 17, Henikoff teaches a method wherein the cell is immobilized on a solid surface, optionally wherein the solid surface comprises a bead or wall of a microtiter plate. (Para. 132 ; Para. 150 ) Regarding claim 19, Henikoff teaches a method wherein the first and/or second DNA molecule further comprises a barcode, a sequencing adaptor, and/or a universal priming site. (Para. 152 -Para.154; Para. 157) Regarding claim 22, Henikoff teaches a method wherein the at least one transposase comprises a Tn5 transposase. (Para. 158) Regarding claim 23, Henikoff teaches a method wherein activating the transposase under low ionic conditions comprises contacting the transposase with Mg ++ , optionally with about 0.1 mM Mg ++ to about 10 mM Mg ++ . (Para. 155 ; Para. 223 ) Regarding claim 24, Ref teaches a method wherein the at least one transposase comprises a Mu transposase, an IS5 transposase, or an IS91 transposase. (Para. 159) Regarding claim 26, Henikoff teaches a method wherein the least one transposome comprises at least two different transposases, and wherein the different transposases integrate different DNA sequences into the chromatin DNA. (Para. 130) Regarding claim 27, Henikoff teaches a method wherein the method is performed with a plurality of first affinity reagents, thereby producing a plurality of excised tagged DNA segments, and wherein the method further comprises isolating a plurality of excised tagged DNA segments, and optionally further comprising analyzing the isolated tagged DNA segments, optionally wherein analyzing the isolated tagged DNA segments comprises determining the nucleotide sequence of the tagged DNA segments, optionally using sequencing or hybridization techniques with or without amplification. Regarding claim 34, Henikoff teaches a method wherein the cell and/or the nucleus of the cell is permeabilized by contacting the cell with digitonin. (Para. 29; Para. 130) Regarding claim 35, Henikoff teaches a method further comprising subjecting the excised DNA to salt fractionation. (Para. 134; Para. 318) Regarding claim 36, Henikoff teaches a method wherein the NDR marker is a histone modification, optionally methylated H3K4, optionally wherein methylated H3K4 is bi-methylated or tri-methylated, optionally wherein the NDR marker is an initiating form of RNA Polymerase II, optionally serine 5-phosphorylated RNA Polymerase II (RNAPIIS5P) or serine 2-phosphorylated RNA Polymerase II (RNAPIIS2P). (Para. 4; Para. 36) Regarding claim 38, Henikoff teaches a method further comprising contacting the permeabilized cell with a known amount of spike-in DNA configured to facilitate calibration, optionally wherein the spike-in DNA is or comprises exogenous DNA, exogenous chromatin, or recombinant nucleosomes. (Para. 221) Regarding claim 40, Henikoff teaches a method wherein the first affinity reagent is coupled to a plurality of transposomes , a fraction of the plurality of transposomes comprising a known amount of spike-in DNA, and wherein the spike-in DNA can be used for calibration. (Para. 887) Regarding claim 41, Henikoff teaches a method wherein the at least one transposome comprises a fusion protein comprising a first domain comprising a Tn5 transposase domain and second domain comprising a protein A domain, a protein G domain, or a protein A/G hybrid domain. (Para. 67) Regarding claim 42-43, Henikoff teaches a method wherein the method is performed for a plurality of cells and the method further comprises mapping the determined sequences of one or more excised tagged DNA segments to a consensus genome of the plurality of the cells; and further comprising mapping the determined sequence of the excised tagged DNA segment to the genome of the cell. (Para. 223) Regarding claim 44, Henikoff teaches a method wherein the method is performed for a plurality of cells, wherein the excised tagged DNA segments of each of the plurality of cells is tagged with a cell-specific barcode or combination of barcodes that is unique to each cell, optionally wherein the method further comprises application of combinatorial indexing to provide the cell-specific barcode or combination of barcodes to the excised tagged DNA segments of each of the plurality of cells and/or wherein the plurality of cells is disposed in a three-dimensional arrangement and the cell- specific barcode or combination of barcodes is unique to a location in the three-dimensional arrangement, optionally wherein the three-dimensional arrangement is a tissue slice or tissue culture array. (Para. 152) Regarding claim 51, Henikoff teaches a method for preparing a library of excised chromatin DNA comprising the method of claim 1. (Para. 6; Para. 223) Claim Rejections - 35 USC § 103 The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action: A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made. Claim s 1-5, 8-10, 13-15, 17, 19, 22-24, 26-27, 34-36, 38, 40-44 and 51 are rejected under 35 U.S.C. 103 as being unpatentable over Chen et al. (“Chen”; Patent App. Pub. US 20180335424 A1, Nov. 11, 2018) . Interpretation: The limitation “optionally”, recited in claims 10, 15, 17, 23, 27, 36, 38, and 44, is interpreted as rendering the phrase thereafter as an optional limitation to the claim. Chen discloses “The present application provides a method of analyzing protein binding sequence, a method of making sequencing library, and compositions for performing such methods, which employs a transposome complex.” (Abstract). Regarding claim 1, Chen teaches a method wherein “The present invention can be used to study every aspect of epigenetics and epigenomics, such as chromatin accessibility ( or nucleosome occupancy), nucleosome positioning, transcription factor (TF) occupancy, histone modification pattern, etc. ” (Para. 187). Chen teaches a method wherein “ the cell sample can be permeabilized ” (Para. 258 ). Chen teaches a method wherein “ providing a moiety, such as biotin, as an affinity tag for attaching to another molecule which is in solution or bound to a surface. ” (Para. 91 ). Chen teaches a method wherein “ The present invention provides methods of analyzing binding sequences on a chromatin to which a protein of interest binds, methods of sequencing a nucleic acid sequence on a chromosome, methods of making sequencing library, methods of tagging nucleic acids, methods of tagmenting nucleic acids using Tn5 transposome complex, and compositions for performing such methods ” (Para. 7 ). Chen further teaches a method wherein “ a method of analyzing the binding sequences on a chromosome to which a protein of interest binds, comprising: (a) … a plurality of transposon end compositions comprising transposon end described herein into the chromatin or double-stranded nucleic acid fragments thereof (e.g., chromatin DNA) in the presence of a transposase (e.g., Tn5); (b) subjecting the double-stranded nucleic acid fragments inserted with transposon end compositions comprising transposon end to immunoprecipitation using a binding agent (e.g., an antibody or fragments thereof) specifically recognizing the protein of interest (e.g., for use in next-generation sequencing); and (c) analyzing the nucleic acid fragment sequences to which the protein of interest binds. ” (Para. 177 ). Chen teaches a method wherein “incubating the chromatin (native or cross-linked) in an in vitro or in vivo transposition reaction with at least one transposase and a transposon end composition with which the transposase forms a transposome complex …. thereby fragmenting the chromatin (or chromatin DNA) and generating a population of annealed 5'-tagged chromatin (or chromatin DNA) fragments, each of which has the transposon end composition tag on the 5'-end. ” (Para. 177). Chen teaches a method wherein “chromatin (or chromatin DNA) is immunoprecipitated with a binding agent (e.g., an antibody or fragments thereof) specifically recognizing the protein of interest” (Para. 177). Thus, Chen teaches an in situ method for detecting a site of DNA accessibility in the chromatin of a cell, comprising: contacting a permeabilized cell with a first affinity reagent that specifically binds a nucleosome depleted region (NDR) marker, wherein the first affinity reagent is coupled to at least one transposome comprising: at least one transposase; and a transposon comprising: a first DNA molecule comprising a first transposase recognition site; and a second DNA molecule comprising a second transposase recognition site; activating the at least one transposase under low ionic conditions, thereby cleaving and tagging chromatin DNA with the first and second DNA molecules and excising a tagged DNA segment associated with the NDR marker; isolating the excised tagged DNA segment; and determining the nucleotide sequence of the excised tagged DNA segment, thereby detecting the site of DNA accessibility in the chromatin of the cell. The teachings of Chen are documented above in the rejection of claim 1 under 35 U.S.C. 103. Claim s 2, 4, 9, 13, 15, 17, 19, 22, 24, 26-27, 34-36, 38, 41-44, 48 and 51 depend on claim 1 . Claim 3 depends on claim 2, which depends on claim 1. Claim 3 depends on claim 2, which depends on claim 1. Claim 8 depends on claim 5. Claims 5-7 depend on claim 4, which depend on claim 1. Claim 10 depend on claim 9, which depends on claim 1. Claim 14 depends on claim 13, which depends on claim 1. Claim 23 depends on claim 22, which depends on claim 1. Claim 40 depends on claim 38, which depends on claim 1. Regarding claim 2 -3 , Chen teaches a method wherein “ the transposase can further comprise an affinity tag ” (Para. 262 ). Thus, Chen teaches a method wherein the first affinity reagent is directly coupled to at least one transposase; and wherein the first affinity reagent and transposase are disposed in a fusion protein . Regarding claim 4, Chen teaches a method wherein “ (a) immunoprecipitating the chromatin using a binding agent (e.g., an antibody or fragments thereof) specifically recognizing the protein of interest; (b) randomly inserting a plurality of transposon end compositions comprising transposon end described herein into the chromatin or double-stranded nucleic acid fragments thereof (e.g., chromatin DNA) in the presence of a transposase (e.g., Tn5 ); (c) analyzing the nucleic acid fragment sequences to which the protein of interest binds. In some embodiments, the chromatin sample is cross-linked. ” (Para. 178 ). Thus, Chen teaches a method wherein the first affinity reagent is indirectly coupled to the at least one transposase. Regarding claim 5, Chen teaches a method wherein “affinity binding molecule that is bound to a detectable moiety or that can form an affinity binding pair or a specific binding pair with another detectable affinity-binding molecule” (Para. 288) and “ multiple-component affinity tag complexes include ligands and their receptors ” (Para. 285 ). Thus, Chen teaches a method wherein the transposase is linked to a specific binding agent that specifically binds the first affinity reagent. Regarding claim 8, Chen teaches a method wherein “the immunoprecipitating binding agent, in particular the antibody or chemical substance, can be immobilized on surfaces via affinity interactions... Where the immunoprecipitating binding agent is an antibody (or fragments thereof), the Fc-part of the antibody can bind to the surface of the beads via Protein A, Protein G ” (Para. 294 ). Thus, Chen teaches a method wherein the specific binding agent comprises protein A or protein G that specifically binds the first affinity reagent, the second affinity reagent and/or the third affinity reagent. Regarding claim 9 -10 , Chen teaches a method wherein “ In some embodiments, the immunoprecipitating binding agent is an antibody. Such antibodies can specifically recognize any of the protein of interest described herein… In some embodiments, the antibody is an antigen-binding fragment, e.g., a Fab, a Fab′, a F(ab′)2, an Fv fragment, a disulfide stabilized Fv fragment ( dsFv ), or a single-chain Fv ( scFv ). Any antibody or antigen-binding fragment can be used in the present invention, as long as chromatin bound by the antibody or antigen-binding fragment can be recovered ” (Para. 291 ). Thus, Chen teaches a method wherein the first, second, and/or third affinity reagents independently is or comprises an antibody, an antibody- like molecule, a DARPin , an aptamer, a chromatin-binding protein, other specifically binding molecule, or a functional antigen-binding domain thereof; and wherein the antibody-like molecule is an antibody fragment and/or antibody derivative, optionally a single-chain antibody, a bispecific antibody, an Fab fragment, an F(ab)2fragment, a VHH fragment, a VNAR fragment, or a nanobody, optionally wherein the single-chain antibody is a single chain variable fragment ( scFv ), or a single-chain Fab fragment ( scFab ). Regarding claim 13 -14 , Chen teaches a method wherein “ endogenous Mg 2+ ” (Para. 261). Chen teaches a method wherein “facilitated and/or triggered by addition of one or more cations. The cations can be divalent cations such as, for example, Ca2+, Mg2+ and Mn2+.” (Para. 259). Chen teaches a method wherein 10 mM MgCl 2 or less is present in shearing buffers described (Para. 524; Para 527; Para. 532) . Chen teaches a method wherein “ Cell pellets were washed with Shearing buffer … Then the Shearing buffer was diluted 5 times or 10 times ” (Para. 526). Thus, Chen teaches a method wherein the low ionic conditions are characterized by monovalent ionic concentration of less than about 10 mM; and wherein the low ionic conditions are obtained by diluting liquid conditions of the transposase with a Mg ++ solution, removing liquid supernatant from the transposase and replacing it with a low ionic strength solution, and/or conducting a stringent (e.g., 300 mM) wash followed by adding a low ionic strength solution. Regarding claim 15, Chen teaches a method wherein “ Dimethyl pimelimidate (DMP) ” (Para. 294 ). Chen teaches a method wherein “Spermidine” in the TA Reaction buffer (see Materials) (Para. 510). Thus, Chen teaches a method further comprising contacting the permeabilized cell with a polar compound prior to or during the step of activating the transposase under low ionic conditions Regarding claim 17, Chen teaches a method wherein “ fixation of cells ” (Para. 175 ) and “fixed cells or tissue sections” (Para. 324) . Thus, Chen teaches a method wherein the cell is immobilized on a solid surface, optionally wherein the solid surface comprises a bead or wall of a microtiter plate. Regarding claim 19, Chen teaches a method wherein “ ligation of adaptors ” and “barcode sequences to multiplex sequencing experiments ” (Para. 175 ). Thus, Chen teaches a method wherein the first and/or second DNA molecule further comprises a barcode, a sequencing adaptor, and/or a universal priming site. Regarding claim 22, Chen teaches a method wherein “ the transposase is Tn5 ” (Para. 13 ). Thus, Chen teaches a method wherein the at least one transposase comprises a Tn5 transposase. Regarding claim 23, Chen teaches a method wherein “ In general, a suitable in vitro transposition system for use in the methods of the present invention requires, at a minimum, a transposase enzyme of sufficient purity, sufficient concentration, and sufficient in vitro transposition activity ” (Para. 255 ). Chen teaches a method wherein “both prokaryotic (Gram negative and Gram positive bacteria) and eukaryotic (yeast, trypanosome, and mice) where in the presence of endogenous Mg2+, transposon insertion has shown to be random and stable.” (Para. 261) Thus, Chen teaches a method wherein activating the transposase under low ionic conditions comprises contacting the transposase with Mg++, optionally with about 0.1 mM Mg++ to about 10 mM Mg++. Regarding claim 24, Chen teaches a method wherein “ The transposase can be prokaryotic or eukaryotic. Examples of transposase include, but are not limited to, a Tn transposase (e.g. … a MuA transposase …IS5… or any transposase related to and/or derived from those listed above. ” (Para. 259 ). Thus, Chen teaches a method wherein the at least one transposase comprises a Mu transposase, an IS5 transposase, or an IS91 transposase. Regarding claim 26, Chen teaches a method wherein “incubating the chromatin (native or cross-linked) in an in vitro or in vivo transposition reaction with at least one transposase and a transposon end composition with which the transposase forms a transposome complex, the transposon end composition comprising ( i ) a transferred strand that exhibits a transferred transposon end sequence and, optionally, an additional sequence (e.g., sample index tag, restriction site tag, amplification tag) 5′-of the transferred transposon end sequence, and (ii) a non-transferred strand that exhibits a sequence that is complementary to the transferred transposon end sequence, under conditions and for sufficient time wherein multiple insertions into the target DNA occur” (Para 177). Chen teaches a method wherein “the transposase is Tn5” (Para. 13). “Tn5” reads on a transposase that functions as a dimer, thus two different transposases. Thus, Chen teaches a method wherein the least one transposome comprises at least two different transposases, and wherein the different transposases integrate different DNA sequences into the chromatin DNA. Regarding claim 27, Chen teaches a method wherein “inserting a plurality of transposon end compositions comprising transposon end described herein into the chromatin or double-stranded nucleic acid fragments thereof (e.g., chromatin DNA) in the presence of a transposase (e.g., Tn5); (b) subjecting the double-stranded nucleic acid fragments inserted with transposon end compositions comprising transposon end to immunoprecipitation using a binding agent” and “after isolating the 5′-transposon end composition tagged chromatin (or chromatin DNA) captured by immunoprecipitating binding agent, and before reverse-crosslinking and denaturing, the captured 5′-tagged chromatin (or chromatin DNA) is subjected to an additional exonuclease (e.g., Exo III) digestion step to digest the tagged chromatin DNA up to the binding boundary of the protein of interest” (Para. 177). Thus, Chen teaches a method wherein the method is performed with a plurality of first affinity reagents, thereby producing a plurality of excised tagged DNA segments, and wherein the method further comprises isolating a plurality of excised tagged DNA segments, and optionally further comprising analyzing the isolated tagged DNA segments, optionally wherein analyzing the isolated tagged DNA segments comprises determining the nucleotide sequence of the tagged DNA segments, optionally using sequencing or hybridization techniques with or without amplification. Regarding claim 34, Chen teaches a method wherein “ permeabilization agents include … digitonin ” (Para. 258 ). Thus, Chen teaches a method wherein the cell and/or the nucleus of the cell is permeabilized by contacting the cell with digitonin. Regarding claim 35, Chen teaches a method wherein “ addition and subsequent removal of buffered aqueous solutions containing chemicals including salt and detergents. ” (Para. 294 ). Chen teaches a method wherein “Buffer conditions that may alter stringent conditions during cell lysis and isolation may include pH and salt concentration” (Para. 221). Chen teaches a method wherein “to retain integrity of the sample comprising nucleic acid. In addition to heating, … salt … may be added to remove crosslinks” (Para. 299) . Thus, Chen teaches a method further comprising subjecting the excised DNA to salt fractionation. Regarding claim 36, Chen teaches a method wherein “ In some embodiments, the protein of interest is selected from the group consisting of transcription factor, histone, histone modification, chromatin remodeler, chromatin modifier, transcription machinery elements ... ” (Para. 8 ). Thus, Chen teaches a method wherein the NDR marker is a histone modification, optionally methylated H3K4, optionally wherein methylated H3K4 is bi-methylated or tri-methylated, optionally wherein the NDR marker is an initiating form of RNA Polymerase II, optionally serine 5-phosphorylated RNA Polymerase II (RNAPIIS5P) or serine 2-phosphorylated RNA Polymerase II (RNAPIIS2P). Regarding claim 38, Chen teaches a method “using chromatin from another species as spike-in control” (Para. 5). Thus , Chen teaches a method further comprising contacting the permeabilized cell with a known amount of spike-in DNA configured to facilitate calibration, optionally wherein the spike-in DNA is or comprises exogenous DNA, exogenous chromatin, or recombinant nucleosomes. Regarding claim 40, Chen teaches a method wherein “ using chromatin from another species as spike-in control ” (Para. 5 ). Chen teaches a method wherein “combining multiple samples into one experimental reaction, so that variances among different samples during different experiments are significantly minimized, samples can start with smaller amount of cells or nuclei, samples can be quantitatively and qualitatively compared with each other, while different samples can be told apart” Thus, Chen teaches a method wherein the first affinity reagent is coupled to a plurality of transposomes , a fraction of the plurality of transposomes comprising a known amount of spike-in DNA, and wherein the spike-in DNA can be used for calibration. Regarding claim 41, Chen teaches a method wherein “ … in the presence of a transposase (e.g., Tn5); (b) subjecting the double-stranded nucleic acid fragments inserted with transposon end compositions comprising transposon end to immunoprecipitation using a binding agent (e.g., an antibody or fragments thereof) …In some embodiments, the method comprises: incubating the chromatin (native or cross-linked) in an in vitro or in vivo transposition reaction with at least one transposase and a transposon end composition with which the transposase forms a transposome complex… ” (Para. 177 ). Chen teaches a method wherein “the immunoprecipitating binding agent, in particular the antibody or chemical substance, can be immobilized on surfaces via affinity interactions... Where the immunoprecipitating binding agent is an antibody (or fragments thereof), the Fc-part of the antibody can bind to the surface of the beads via Protein A, Protein G” (Para. 294). Thus, Chen teaches a method wherein the at least one transposome comprises a fusion protein comprising a first domain comprising a Tn5 transposase domain and second domain comprising a protein A domain, a protein G domain, or a protein A/G hybrid domain. Regarding claim 42 -43 , Chen teaches a method wherein “The population of cells used in the assay may be composed of any number of cells” (Para. 215). Chen teaches a method wherein “a method of sequencing a nucleic acid sequence on a chromosome, comprising: (a) randomly inserting a plurality of transposon end compositions comprising transposon end … in the presence of a transposase (e.g., Tn5), wherein the transposon end composition comprises, from 5′ to 3′: a sample index tag, an amplification tag, a restriction site tag, and a transposon end; and (b) determining the nucleic acid fragment sequences. ” (Para. 181). Chen also teaches a method wherein “ Based on matching the sequenced nucleic acids to genomic sequences ” (Para. 345 ). Thus, Chen teaches a method wherein the method is performed for a plurality of cells and the method further comprises mapping the determined sequences of one or more excised tagged DNA segments to a consensus genome of the plurality of the cells; and further comprising mapping the determined sequence of the excised tagged DNA segment to the genome of the cell. Regarding claim 44, Chen teaches a method wherein “ multiple barcoded biological samples ” (Para. 83 ). Thus, Chen teaches a method wherein the method is performed for a plurality of cells, wherein the excised tagged DNA segments of each of the plurality of cells is tagged with a cell-specific barcode or combination of barcodes that is unique to each cell, optionally wherein the method further comprises application of combinatorial indexing to provide the cell-specific barcode or combination of barcodes to the excised tagged DNA segments of each of the plurality of cells and/or wherein the plurality of cells is disposed in a three-dimensional arrangement and the cell- specific barcode or combination of barcodes is unique to a location in the three-dimensional arrangement, optionally wherein the three-dimensional arrangement is a tissue slice or tissue culture array. Regarding claim 51, Chen teaches a method wherein “ a method of preparing sequencing library starting from ChIPed DNA, input DNA, dsDNA, or any nucleic acid ” (Para. 180 ). Thus, Chen teaches a method for preparing a library of excised chromatin DNA comprising the method of claim 1. Therefore, the invention as recited in claims 1-5, 8-10, 13-15, 17, 19, 22-24, 26-27, 34-36, 38, 40-44 and 51 are prima facie obvious over the prior art Chen et al . One of ordinary skill in the art would have had a reasonable expectation of success given the lack of novelty. It would have been obvious to provide an in -situ method for detecting a site of DNA accessibility in the chromatic of a cell, according to the limitations of the instant application claims 1-5, 8-10, 13-15, 17, 19, 22-24, 26-27, 34-36, 38, 40-44 and 51 based on Chen et al. (Patent App. Pub. No. US 20180335424 A1). Conclusion The prior art made of record and not relied upon is considered pertinent to applicant's disclosure: Kaya-Okur et al. (2019). CUT&Tag for efficient epigenomic profiling of small samples and single cells. Nature communications, 10(1), 1930. (pg.8-9; entire document) Claim 1. 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