Abstract: Selecting which sub-sequences in a database of nucleic acid such as 16S rRNA are highly characteristic of particular groupings of bacteria, microorganisms, fungi, etc. on a substantially phylogenetic tree. Also applicable to viruses comprising viral genomic RNA or DNA. A catalogue of highly characteristic sequences identified by this method is assembled to establish the genetic identity of an unknown organism. The characteristic sequences are used to design nucleic acid hybridization probes that include the characteristic sequence or its complement, or are derived from one or more characteristic sequences. A plurality of these characteristic sequences is used in hybridization to determine the phylogenetic tree position of the organism(s) in a sample. Those target organisms represented in the original sequence database and sufficient characteristic sequences can identify to the species or subspecies level. Oligonucleotide arrays of many probes are especially preferred.
Type:
Grant
Filed:
January 26, 2002
Date of Patent:
July 3, 2012
Assignee:
Technology Licensing Co. LLC
Inventors:
George E. Fox, Richard C. Willson, III, Zhengdong Zhang
Abstract: The method is used for separating nucleic acids and other similar constructs. It involves selective introduction, enhancement, or stabilization of affinity handles such as single-strandedness in the undesired (or desired) nucleic acids as compared to the usual structure (e.g., double-strandedness) of the desired (or undesired) nucleic acids. The undesired (or desired) nucleic acids are separated from the desired (or undesired) nucleic acids due to capture by methods including but not limited to immobilized metal affinity chromatography, immobilized single-stranded DNA binding (SSB) protein, and immobilized oligonucleotides.
Type:
Grant
Filed:
December 16, 2003
Date of Patent:
February 22, 2011
Assignees:
University of Houston, Technology Licensing Co. LLC
Inventors:
Richard Coale Willson, III, Luis Antonio Cano
Abstract: An immobilized metal affinity chromatography (IMAC) method for separating and/or purifying compounds containing a non-shielded purine or pyrimidine moiety or group such as nucleic acid, presumably through interaction with the abundant aromatic nitrogen atoms in the purine or pyrimidine moiety. The method can also be used to purify compounds containing purine or pyrimidine moieties where the purine and pyrimidine moieties are shielded from interaction with the column matrix from compounds containing a non-shielded purine or pyrimidine moiety or group. Thus, double-stranded plasmid and genomic DNA, which has no low binding affinity can be easily separated from RNA and/or oligonucleotides which bind strongly to metal-charged chelating matrices. IMAC columns clarify plasmid DNA from bacterial alkaline lysates, purify a ribozyme, and remove primers and other contaminants from PCR reactions. The metal ion affinity of yeast RNA decreases in the order: copper (II), nickel (II), zinc (II), and cobalt (II).
Abstract: RNA, preferably messenger RNA, is purified by use of selective precipitation, preferably by addition of compaction agents. Also included is a scalable method for the liquid-phase separation of DNA from RNA and RNA may also be recovered by fractional precipitation. Specific classes of compounds e.g. phase transfer catalysts (PTCs), most preferably selected polyamines of U.S. Pat. No. 6,617,108 polyamines which are quaternary compounds are unexpectedly potent in causing selective precipitation of DNA away from RNA, at low concentrations and in the presence of relatively elevated ionic strength selective removal of DNA can also remove both RNA and DNA, leaving behind a mixture which is advantageous for the further purification of, e.g., proteins.
Type:
Grant
Filed:
February 1, 2006
Date of Patent:
August 4, 2009
Assignee:
Technology Licensing Co. LLC
Inventors:
Richard C. Willson, III, Richard Don Goodin
Abstract: Preferred embodiments of the invention include purification of DNA, preferably plasmid DNA, by use of selective precipitation, preferably by addition of compaction agents. Also, included is a sealable method for the liquid phase separation of DNA from RNA. RNA may also be recovered by fractional precipitation according to the invention. Applicants have discovered that RNA, commonly the major contaminant in DNA preparations, can be left in solution while valuable purified plasmid DNA is directly precipitated. Additional aspects of the invention include mini-preps, preferably of plasmid and chromosomal DNA, to obtain sequenceable and restriction digestible DNA in high yields in multiple simultaneous procedures. Still further aspects disclose enhanced stripping of the compaction agent by a stripping method comprising high salt addition and pH shift, and combinations of these techniques. Also, disclosed is a method of assay in which a labeled probe is precipitated when it is hybridized to a target, (e.g.
Abstract: A method for treatment of a disease comprising vasospasm or other symptom alleviable by smooth muscle relaxation and a vasodilator delivery system. The figure is a TCD of MCA post nitroglycerine spray obtained during continuous monitoring.
Abstract: An improved manganese and/or chromium-promoted catalytic process, catalyst and method of manufacture for heavy hydrocarbon conversion, optionally in the presence of nickel and vanadium on the catalyst and in the feed stock, to produce lighter molecular weight fractions, including lower olefins and higher isobutane than normally produced. This process is based on the discovery that two "magnetic hook" elements, namely manganese and chromium, previously employed as magnetic enhancement agents to facilitate removal of old catalyst, or to selectively retain expensive catalysts, can also themselves function as selective cracking catalysts, particularly when operating on feeds containing significant amounts of nickel and vanadium, and especially where economics require operating with high nickel- and vanadium-contaminated and containing catalysts.