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Denaturing in Biological Technology

Separation of HLA-DRB Alleles by Denaturing Gradient Gel Electrophoresis using the DCode System

L. A. Knapp, 1, 2 E. Lehmann, 2 L. Hennes, 3 M. E. Eberle, 2 and D. I. Watkins 2, 3 1 Department of Biological Anthropology, University of Cambridge,U.K. 2 Wisconsin Regional Primate Research Center, University of Wisconsin, ...

Microbial Diversity in Ground and Surface Water Analyzed by Denaturing Gradient Gel Electrophoresis Using the DCode System

Bernd Eschweiler and Beate Kilb, Institute for Water Research, Schwerte, Germany Introduction Bacterial diversity in environmental samples is usually determined by a characterization of isolated strains. A problem for the analysis and ...

Identification of Disease Causing Mutations in Phenylketonuria by Denaturing Gradient Gel Electrophoresis Using the DCode System

Luc Michiels 1 , Baudouin Franois 1, 2 , and Jef Raus 1, 2 , 1 Dr. L. Willems-Instituut and 2 Limburgs Universitair Centrum, Universitaire Campus, B-3590 Diepenbeek, Belgium Introduction Phenylketonuria (PKU) is a hereditary...

Detection of p53 Gene in Breast Cancer by Denaturing Gradient Gel Electrophoresis and the DCode System

Akira Ichikawa and Seiji Igarashi Tochigi Cancer Center, Tochigi, Japan Introduction Denaturing gradient gel electrophoresis (DGGE) is one of the most consistently used mutation scanning methods. It has evolved enormously o...

Detection of Variation in Highly Polymorphic Mhc Genes by Denaturing Gradient Gel Electrophoresis Using the DCode System

Kristi M. Miller, Tobi J. Ming and Ruth E. Withler, Pacific Biological Station, Department of Fisheries and Oceans, Nanaimo, B.C. V9R 5K6, Canada Introduction We describe a rapid and sensitive method for the detection of nucleotide s...

Detection of Mutations in the Fas Antigen of Lymphoma Tumors by RT-PCR and Denaturing Gradient Gel Electrophoresis

Terry H. Landowski, Ibrahim Buyuksal, and William S. Dalton H. Lee Moffitt Cancer Center and Research Institute, University of South Florida, 12902 Magnolia Drive, Tampa FL 33612 Introduction Identifying of mutations within specific genes has...

Detection of K-ras Point Mutations in the Pancreas by Constant Denaturing Gel Electrophoresis Using the DCode System

Martin A.O.H. Menke*, Axel Reinecke-Lthge*, Barbara Mllmann*, Angela Kellner, Jutta Lttges*, Gnter Klppel* * Department of Pathology and Department of Hematopathology, Christian- Albrechts University Kiel, Michaelisstrae 11, D-24105 Kiel, Germany. ...

A Multiple Mutation Model System as a Test Development and Training Tool for Denaturing Gradient Gel Electrophoresis

Michael Hepburn and Glenn A. Miller, Ph.D., Molecular Profiling Laboratory, Genzyme, Framingham, MA Introduction The search for sequence variations in genomic DNA is important in the study of genes that play a role in the devel...

Transgenomic Completes Licensing Option With Dana-Farber Cancer Institute on Cold-PCR for Enrichment of DNA Mutations

...the same gene are mixed and re-annealed, variations between these sequences cause double-stranded DNA heteroduplexes to form. The WAVE System employs denaturing HPLC to separate these homo- and hetero-duplexes by ion-pairing reverse-phase HPLC. This technology has been in widespread use for genomic analysis be...

Noridian Medicare Now Covers Renessa(R) Treatment for Incontinence in Women

...atural opening of the urethra (transurethral). The probe heats multiple small treatment sites in the submucosa of the bladder neck and upper urethra, denaturing the patient's own collagen in the tissue, thereby reducing or eliminating leaks. The Renessa treatment can be performed in the convenience of a physi...

Novasys Medical Secures $49.5 Million in Series D Financing to Expand Commercialization of Its Renessa(R) Treatment for Incontinence in Women

...natural opening of the urethra (transurethral). The probe heats multiple small treatment sites in the submucosa of the bladder neck and upper urethra, denaturing collagen in the tissue, thereby reducing or eliminating leaks. The Renessa treatment can be performed in the convenience of a physician's office using...

Americas Third Coast nurtures biotechnology

...n, can harm a proteins structure and activity. The novel, detergent-based YeastBuster, BugBuster, and CytoBuster Reagents perforate cell walls without denaturing proteins, continues Harris. After a brief incubation, target cells are harvested by centrifugation and the protein is extracted. Novagen also mark...

Denaturing HPLC (DHPLC)

...ed population is analysed by HPLC under partially denaturing temperatures, the heteroduplexes elute from the c...e required temperature. Optimisation of the denaturing temperature can be performed by trial or error. ...d increase as the temperature is increased. At the denaturing temperature, the retention time will decrease wit...

Protein Depletion for Plasma and Serum Proteomic Analysis

... Stecher G, Bonn GK, Huber LA: Enrichment of low-abundant serum proteins by albumin/immunoglobulin G immunoaffinity depletion under partly denaturing conditions. Electrophoresis 2005, 26 :2843-2849.
  • Lowenthal MS, Mehta AI, Frogale K, Bandle RW, Araujo RP, Hoo...

    New gravity-flow column gives faster and simpler purification of histidine-tagged proteins

    ...ifications can be performed either under native or denaturing conditions, and a number of additives can be used.... Ni-NTA Superflow columns were also compared under denaturing conditions. Histidine-tagged Maltose Binding Prote...by lowering the pH from pH 8.0 to pH 4.5. Although denaturing conditions increase the purification time due to t...

    Use of the DCode System to Detect the Food-Borne Bacterial Pathogen Listeria monocytogenes

    ...bo polymerase was added after the first denaturing step, at a temperature of 80 C, a techn... DGGE DGGE was performed with a DCode denaturing gel electrophoresis system (Bio-Rad Lab...CA). A parallel gel was prepared with a denaturing gradient ranging from 15 to 40% as described in Ap...

    Protein Truncation Tests Using the DCode System

    ...hionine. Protein products were run on a 16 x 20 cm, 14% SDS-PAGE denaturing polyacrylamide gel using the DCode universal mutation detection ...sm (SSCP), conformation-sensitive gel electrophoresis (CSGE) and denaturing gradient gel electrophoresis (DGGE). Protein truncation test ana...

    Detection of Apolipoprotein Gene Variants by DGGE Using the DCode System

    ...e hereditary diseases. We have used the denaturing gradient gel electrophoresis method (DGGE) 1 usin... or 58 C (apoB), in each case for 30 s. The denaturing and elongation steps were 96 C for 30 s... to predict the percentage range of the denaturing gradient to be used in the gels. Multiple aliquots...

    Monitoring Bacterial Genetic Diversity in a Freshwater Lake Using TTGE and DNA Sequence Analysis

    ...d of choice for sequence variation studies. In the commonly used denaturing Gradient Gel Electrophoresis, 5 electrophoretic separation is p...temperature in the presence of a chemical gradient. In TTGE, the denaturing environment is formed by a constant concentration of denaturant ...

    Heteroduplex Analysis Using the DCode System

    ...y adding mutant and wild-type DNA in the same PCR reaction or by denaturing and renaturing a mixture of mutant and wild-type DNA. Both mutan... formed by mixing the mutant and wild-type samples together, then denaturing and renaturing the DNA. The ability to reso...

    Detection of Mutations in the CYP21 Gene Using the DCode System

    ... a complicated matter. We have used the denaturing gradient gel electrophoresis method (DG...ting heteroduplex molecules. denaturing Gradient Gel Electrophoresis denaturing gradient gel electrophoresis was carried out using...

    Analysis of Bacterial Assemblage Genetic Diversity in Environmental Samples Using the DCode System

    ...Molecular techniques like the polymerase chain reaction (PCR)* have enhanced the ability to examine ribosomal genes of bacteria. denaturing gradient gel electrophoresis (DGGE) has proven to be a powerful tool for analyzing PCR-amplified bacterial 16S rDNA fragments. T...

    The Bio-Rad Silver Stain, Rev E

    ...sality Stains gels of various composition (see References) including IEF, 2-D, gradient and peptide gels, and nucleic acids in denaturing gels Adjunct to Protein Identification and Characterization Protein-specific slopes (OD vs. protein concentration plot) differe...

    Performance Comparison of the Experion Automated Electrophoresis System and a Competing Automated System for Protein Analysis, Rev A

    ...lysis of protein samples. The Experion system uses the Experion Pro260 analysis kit for the separation of 10260 kD proteins under denaturing conditions. Typically, a Pro260 separation and analysis of up to ten samples is accomplished in approximately 30 min. This autom...

    Using the DCode System to Identify DNA Sequence Variation for Studies of Population Structure in Marine Organisms

    ...enetic change in wild or domesticated populations. denaturing gradient gel electrophoresis using the ...og PCR products, perpendicular gradient denaturing gels were run to determine their melting ... samples. Although identification of haplotypes by denaturing gel electrophoresis does not yield the ...

    Real-Time Quantification of Genomic DNA Using DyNAzyme II DNA Polymerase and SYBR Green I Dye

    ...rk with SYBR Green I. Because SYBR Green I stabilizes dsDNA, the denaturing temperature of the cycling protocol should be raised approximate...of read temperatures, as well as the guidelines for DMSO use and denaturing and annealing temperatures, provides the user with some basic to...

    Temporal Temperature Gradient Electrophoresis: A Powerful Technique to Screen Mutations

    ...ngle stranded conformation polymorphism (SSCP), 1 denaturing gradient gel electrophoresis (DGGE), 2 constant denaturing gel electrophoresis (CDGE), 3 temperat...re gradient. 7,8 In TTGE, the denaturing environment is formed by a constant concentration ...

    TTGE on the DCode System

    ...TGE) exploits the principles upon which denaturing gradient gel electrophoresis (DGGE) and constant denaturing gel electrophoresis (CDGE) are based. TTGE combine...in TTGE there is no need for a chemical denaturing gradient which is the advantage in CDGE...

    Separation of 16S rRNA PCR Products From a Model Community Using Temporal Temperature Gradient Gel Electrophoresis, Rev A

    ... (TTGE). In the commonly used denaturing Gradient Gel Electrophoresis, 6 electrophoretic ...ce of a chemical gradient. In TTGE, the denaturing environment is formed by a constant con... of Bacteria in a Biodegraded Wall Painting by denaturing Gradient Gel Electrophoresis of PCR-Amp...

    Detection of Mutant K-ras in a Kindred With Hereditary Pancreatic Cancer by DGGE

    ...ciated with development of pancreatic insufficiency prior to the diagnosis of cancer. 2 We have utilized the DCode system for denaturing gradient gel electrophoresis (DGGE) to identify patient samples with K-ras mutations. Materials and Methods ...

    DGGE Analysis for Non-Invasive Studies of Primate Diet

    ....edu Introduction The applications of denaturing gradient gel electrophoresis (DGGE) to the investigation and dia...ture of PCR products, they were first analyzed on a perpendicular denaturing gradient gel poured with Bio-Rads Model 475 Gradient Former. The ...

    Continuous-Elution Electrophoresis for Purification of the Baculovirus-Expressed Coronavirus Structural Proteins, Rev A

    ...o a 4% (1.5 cm) polyacrylamide stacking gel with a 10% (8 cm) polyacrylamide separating gel. Proteins were electrophoresed under denaturing conditions at constant voltage (200 V) for 3.5 h with a flow rate of 0.05 ml/min. After 3.5 h, fractions were collected in 5 min...

    DGGE Analysis Using the DCode System, Rev B

    ... Introduction denaturing gradient gel electrophoresis (DGGE) is one of seve...hanges or point mutations. In DGGE, the denaturing environment is created by a combination ...ists of 7 M urea and 40% formamide. The denaturing gradient may be formed perpendicular or...

    Purification of Insulin on UNO S1 Column, Rev A

    ... on the market. Although insulin is well characterized and understood, it is often difficult to purify because of the harsh pH and denaturing conditions needed to solubilize and disaggregate the protein. A comparison of the UNO S1 (Bio-Rad catalog # 720-0021) and a popu...

    Nuclease Protection Assays: The Basics

    ...ated. These products are separated on a denaturing polyacrylamide gel and are visualized b...ease digestion, product separation on a denaturing 6% acrylamide gel, and a four-hour exposure ...clease Protection Assay Kit Is Right for Me? " denaturing Acrylamide Gel Electrophoresi...

    Northern Analysis: The Basics

    ...resis in an agarose gel under denaturing conditions. The RNA is then t... RNA) Probe generation denaturing agarose gel electrophoresis Transfer...obes by asymmetric PCR. denaturing Agarose Gel Electrophoresis ...

    Your Data: Silencing Hepatitis C Virus Replication With In Vitro Transcribed siRNAs

    ...r NS3, NS5B, GAPDH, or an irrelevant scrambled (scr) siRNA 2, 4, and 6 days previously. Equal amounts of RNA were separated on a denaturing agarose gel, transferred to membrane and hybridized with probes to HCV and GAPDH. GAPDH signal was used for normalization of the...

    Ultra-Fast Purification of Small Nucleic Acids

    ... only 12 minutes. flashPAGE purification is based on passing up to 10 g nucleic acids through a proprietary denaturing gel matrix, provided as pre-cast gel cartridges, into the lower buffer collection chamber. The lower buffer ...

    The World's Best DNase Improved TURBO DNA-free

    ...DNase inactivation protocols rely on either phenol/chloroform extraction to remove the DNase enzyme, or on a denaturing thermal inactivation step. Both procedures are undesirable. Organic extractions are tedious and prone to RNA...

    Stabilizing RNA in Fresh and Frozen Tissues for RNA Isolation

    ...ed directly from a frozen state or thawed in RNA later -ICE at -20C overnight. (A) shows the ethidium bromide stained RNA in a denaturing agarose gel. (B) shows the results of Northern blot analysis of the same gel hybridized to radiolabeled probes for -actin, GAP...
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