公开(公告)号：US10272409B2

公开(公告)日：2019-04-30

申请号：US12080720

申请日：2008-04-04

申请人： Michael E. Hogan ,  Sergy Lemeshko ,  Yuri Belosludtsev ,  Thomas F. Powdrill ,  Rahul Mitra ,  Joseph G. Utermohlen ,  Frederick H. Eggers

发明人： Michael E. Hogan ,  Sergy Lemeshko ,  Yuri Belosludtsev ,  Thomas F. Powdrill ,  Rahul Mitra ,  Joseph G. Utermohlen ,  Frederick H. Eggers

IPC分类号： C12Q1/68 ,  C12P19/34 ,  B01J19/00

摘要： Provided herein are biomolecular hybridization devices comprising a substrate with a permanently and covalently attached surface of functional groups and an adsorbed monolayer of unmodified, single-stranded oligonucleotides all of which are 10 to about 24 bases in length as a saturated film of constrained oligonucleotides on the surface via direct non-covalent phosphate-surface adsorptive contact of substantially all phosphate groups of each oligonucleotide. The constrained oligonucleotides are effective to dissociably hybridize to a complementary single-stranded nucleic acid with asymmetric, non-helical base pairing and without oligonucleotide dissociation from the surface of the device. Also, provided are methods for hybridizing solution-state target nucleic acids to probe nucleic acids and for identifying a nucleotide sequence to which a nucleotide-binding protein binds using the biomolecular hybridization devices.

公开(公告)号：US07354710B2

公开(公告)日：2008-04-08

申请号：US10193938

申请日：2002-07-11

申请人： Michael Hogan ,  Sergy Lemeshko ,  Yuri Belosludtsev ,  Tom Powdrill ,  Rahul Mitra

发明人： Michael Hogan ,  Sergy Lemeshko ,  Yuri Belosludtsev ,  Tom Powdrill ,  Rahul Mitra

IPC分类号： C12Q1/68

CPC分类号： C12Q1/6837 ,  B01J19/0046 ,  B01J2219/00376 ,  B01J2219/00387 ,  B01J2219/00497 ,  B01J2219/005 ,  B01J2219/00529 ,  B01J2219/00576 ,  B01J2219/00605 ,  B01J2219/00608 ,  B01J2219/00612 ,  B01J2219/00626 ,  B01J2219/00637 ,  B01J2219/00653 ,  B01J2219/00659 ,  B01J2219/00677 ,  B01J2219/00691 ,  B01J2219/00722 ,  C40B60/14 ,  C12Q2523/308

摘要： The present invention relates to simple method to fabricate DNA hybridization devices based upon adsorptive attachment of oligonucleotides to a positively charged surface. Such adsorbed oligonucleotide probes form a densely packed monolayer, which retains capacity for base-pair specific hybridization with a solution state nucleic acid target strand to form the duplex. However, both strand dissociation kinetics and the rate of DNase digestion suggest on symmetry grounds that solution-state nucleic acid binds to such adsorbed oligonucleotides to form a highly asymmetric and unwound duplex, with structural details that are substantially different from that known for the Watson-Crick DNA duplex. This novel nucleic acid duplex form can serve as the basis for a new class of hybridization device and methods for their use. It is also disclosed that new methods of nucleic acid duplex detection can be developed which are based upon the interaction of enzymes and dye labels with the unique structural characteristics of the non-helical duplex described herein. Preferred implementations of the invention include DNA microarrays, bead-based nucleic acid analysis, microelectronic devices to detect nucleic acid hybridization and more traditional methods of laboratory analysis, including hybridization on membranous and other solid supports.

摘要翻译： 本发明涉及基于将寡核苷酸吸附到带正电的表面上的DNA杂交装置的简单方法。 这种吸附的寡核苷酸探针形成密集填充的单层，其保留与溶液状态核酸靶链的碱基对特异性杂交以形成双链体的能力。 然而，链分解动力学和DNA酶消化速率都表明溶液状态核酸与这些吸附的寡核苷酸结合以形成高度不对称和未解链双链体的对称性，结构细节与Watson- 克里克DNA双链体。 这种新的核酸双相形式可以作为新一类杂交装置及其使用方法的基础。 还公开了可以开发新的核酸双相检测方法，其基于酶和染料标记与本文所述的非螺旋双螺旋的独特结构特征的相互作用。 本发明的优选实施方案包括DNA微阵列，基于珠的核酸分析，用于检测核酸杂交的微电子装置和更传统的实验室分析方法，包括膜质和其它固体支持物上的杂交。

公开(公告)号：US20090011949A1

公开(公告)日：2009-01-08

申请号：US12080720

申请日：2008-04-04

申请人： Michael E. Hogan ,  Sergy Lemeshko ,  Yuri Belosludtsev ,  Thomas F. Powdrill ,  Rahul Mitra ,  Joseph G. Utermohlen ,  Frederick H. Eggers

发明人： Michael E. Hogan ,  Sergy Lemeshko ,  Yuri Belosludtsev ,  Thomas F. Powdrill ,  Rahul Mitra ,  Joseph G. Utermohlen ,  Frederick H. Eggers

IPC分类号： C40B40/06 ,  C40B30/04

CPC分类号： B01J19/0046 ,  B01J2219/00376 ,  B01J2219/00497 ,  B01J2219/005 ,  B01J2219/00576 ,  B01J2219/00605 ,  B01J2219/00653 ,  B01J2219/00659 ,  B01J2219/00677 ,  B01J2219/00691 ,  B01J2219/00722

摘要： Provided herein are biomolecular hybridization devices comprising a substrate with a permanently and covalently attached surface of functional groups and an adsorbed monolayer of unmodified, single-stranded oligonucleotides all of which are 10 to about 24 bases in length as a saturated film of constrained oligonucleotides on the surface via direct non-covalent phosphate-surface adsorptive contact of substantially all phosphate groups of each oligonucleotide. The constrained oligonucleotides are effective to dissociably hybridize to a complementary single-stranded nucleic acid with asymmetric, non-helical base pairing and without oligonucleotide dissociation from the surface of the device. Also, provided are methods for hybridizing solution-state target nucleic acids to probe nucleic acids and for identifying a nucleotide sequence to which a nucleotide-binding protein binds using the biomolecular hybridization devices.

摘要翻译： 本文提供的生物分子杂交装置包括具有永久且共价连接的官能团表面的底物和未修饰的单链寡核苷酸的吸附单层，其长度为10至约24个碱基，作为约束寡核苷酸的饱和膜 通过每个寡核苷酸的基本上所有磷酸基团的直接非共价磷酸盐表面吸附接触表面。 受约束的寡核苷酸有效地与具有不对称，非螺旋碱基配对并且没有寡核苷酸与装置表面解离的互补单链核酸解离地杂交。 此外，提供了用于将溶液状态靶核酸杂交以探测核酸并用于鉴定核苷酸结合蛋白使用生物分子杂交装置结​​合的核苷酸序列的方法。