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Patent
LaserGen | Date: 2016-08-08

Provided herein are methods of using photocleavable labels for multiplex and serial antigen detection. The methods comprise detecting the presence of photocleavable labels, which are conjugated through functional linkers to antigen-binding complexes, which in turn non-covalently bind to antigens. The presence of a photocleavable label is indicative of the presence of an antigen specifically or selectively bound by an antigen-binding complex. Also provided are apparatuses for using photocleavable labels for multiplex and serial antigen detection.


Chen X.,University of Wisconsin - Madison | Chen X.,LaserGen | Raggio C.,Hospital for Special Surgery | Campagnola P.J.,University of Wisconsin - Madison
Optics Letters | Year: 2012

We report the use of second-harmonic generation (SHG) microscopy in conjunction with circular dichroism (CD) to differentiate normal skin from that in the connective tissue disorder osteogenesis imperfecta (OI). Osteogenesis imperfecta results from mutations in the collagen triple helix, where the individual chains are defective, leading to abnormal folding, and ultimately, abnormal fibril/fiber organization. Second-harmonic-generation circular dichroism successfully differentiated normal human and OI skin tissues, whereas other SHG polarization schemes did not provide discrimination, suggesting this approach has high sensitivity for studying the difference in chirality in the mutated collagen. We further suggest that the method has clinical diagnostic value, as it could be performed with minimal invasion. © 2012 Optical Society of America.


Metzker M.L.,Baylor College of Medicine | Metzker M.L.,LaserGen
Nature Reviews Genetics | Year: 2010

Demand has never been greater for revolutionary technologies that deliver fast, inexpensive and accurate genome information. This challenge has catalysed the development of next-generation sequencing (NGS) technologies. The inexpensive production of large volumes of sequence data is the primary advantage over conventional methods. Here, I present a technical review of template preparation, sequencing and imaging, genome alignment and assembly approaches, and recent advances in current and near-term commercially available NGS instruments. I also outline the broad range of applications for NGS technologies, in addition to providing guidelines for platform selection to address biological questions of interest. © 2010 Macmillan Publishers Limited. All rights reserved.


The present invention relates generally to 3-OH unblocked nucleotides and nucleosides labeled and unlabeled with 5-methoxy-substituted nitrobenzyl-based photocleavable terminating groups for use in methods and systems related to DNA and RNA sequencing and analysis. These compounds may be used as reversible terminators as they exhibit fast nucleotide incorporation kinetics, single-base termination, high nucleotide selectivity, and rapid terminating group cleavage that results in a naturally occurring nucleotide.


Grant
Agency: Department of Defense | Branch: Defense Health Program | Program: SBIR | Phase: Phase I | Award Amount: 150.00K | Year: 2012

Pathogen detection involving microbial forensics is an emerging field that presents enormous challenges for both the scientific and legal communities. Microbial pathogens of humans represent a highly diverse set of organisms known to cause disease and death. Microbes have also developed a number of elaborate mechanisms for generating natural genetic diversity. One major goal of microbial forensics is to use this genetic diversity to identify the source of a pathogen used to commit a crime or war offensive. Recent advances in next-generation sequencing (NGS) technologies have the potential to significantly alter the technological approaches used in characterizing field samples. This Phase I SBIR grant application proposes three aims: (i) identify the most efficient NGS platform by sequencing E. coli MG1655 using six platforms, (ii), conduct mixing experiments using purified gram negative and gram positive bacteria using the platform selected in aim (i), and (iii) conduct mixing experiments described in aim (ii) in the presence of human blood to simulate animal wound models. The outcome of the Phase I study lays the foundation for a Phase II proposal to develop method/system that would be portable and useful as a point-of-care device for pathogen identification in the field.


Provided are novel nucleotides, nucleoside, and their derivatives described herein, that can be used in DNA sequencing technology and other types of DNA analysis. In one embodiment, the nucleotide or nucleoside with an unprotected 3-OH group is derivatized at the nucleobase to include a fluorescent dye attached via a linker to a photocleavable terminating group. The photocleavable-fluorescent group is designed to terminate DNA synthesis as well as be cleaved so that DNA oligomers can be sequenced efficiently in a parallel format. The design of such rapidly cleavable fluorescent groups on nucleotides and nucleosides can enhance the speed and accuracy of sequencing of large oligomers of DNA in parallel, to allow rapid whole genome sequencing, and the identification of polymorphisms and other valuable genetic information, as well as allowing further manipulation and analysis of nucleic acid molecules in their native state following cleavage of the fluorescent group.


The present invention relates generally to 3-OH unblocked nucleotides and nucleosides labeled and unlabeled with 5-methoxy-substituted nitrobenzyl-based photocleavable terminating groups for use in methods and systems related to DNA and RNA sequencing and analysis. These compounds may be used as reversible terminators as they exhibit fast nucleotide incorporation kinetics, single-base termination, high nucleotide selectivity, and rapid terminating group cleavage that results in a naturally occurring nucleotide.


The present invention relates generally to labeled and unlabled cleavable terminating groups and methods for DNA sequencing and other types of DNA analysis. More particularly, the invention relates in part to nucleotides and nucleosides with chemically cleavable, photocleavable, enzymatically cleavable, or non-photocleavable groups and methods for their use in DNA sequencing and its application in biomedical research.


The present invention relates generally to labeled and unlabeled cleavable terminating groups and methods for DNA sequencing and other types of DNA analysis. More particularly, the invention relates in part to nucleotides and nucleosides with chemically cleavable, photocleavable, enzymatically cleavable, or non-photocleavable groups and methods for their use in DNA sequencing and its application in biomedical research.


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