Profile matching and profile subtraction: application of computer-based image analysis system for two-dimensional electrophoresis gels.

abstract：:Procedures utilizing Chelex 100 chelating resin have been developed for extracting DNA from forensic-type samples for use with the PCR. The procedures are simple, rapid, involve no organic solvents and do not require multiple tube transfers for most types of samples. The extraction of DNA from semen and very small blo...

journal_title：BioTechniques

authors： Walsh PS,Metzger DA,Higuchi R

更新日期：1991-04-01 00:00:00

abstract：:A simple, nondestructive PCR-based screening method has been developed for identifying putative transgenic soft white winter wheat (Triticum aestivum L.) carrying the coat protein gene of wheat streak mosaic virus. Removal of the endosperm end of individual seed provided sufficient material for DNA extraction and PCR....

journal_title：BioTechniques

authors： McCarthy PL,Hansen JL,Zemetra RS,Berger PH

更新日期：2002-03-01 00:00:00

abstract：:Another method has been developed to attach synthetic peptides to solid supports for use in enzyme immunoassays. The method is based on passively adsorbing a synthetic peptide to a solid-phase support, then further attaching more of the same peptide by means of cross-linking to the previously adsorbed peptide. This me...

journal_title：BioTechniques

authors： Leahy DC,Shah DO,Todd JA

更新日期：1992-11-01 00:00:00

abstract：:The study of gene regulation in cardiac myocytes requires a reliable in vitro model. However, monolayer cultures used for this purpose are typically not exposed to electrical stimulation, though this has been shown to strongly affect cardiomyocyte gene expression. Based on pacemakers for clinical use, we developed an ...

journal_title：BioTechniques

authors： Martherus RS,Zeijlemaker VA,Ayoubi TA

更新日期：2010-01-01 00:00:00

abstract：:We describe a method for rapid radioactive labeling of PCR product. The method, employing the Klenow fragment of DNA polymerase I, consumes little product, requires no product purification and takes under 30 minutes. ...

journal_title：BioTechniques

authors： McDaniel TK,Huang Y,Yin J,Needleman SW,Meltzer SJ

更新日期：1991-08-01 00:00:00

abstract：:We describe a multipurpose eukaryotic expression vector that incorporates the following features: restriction sites for in-frame insertion of cDNAs of interest between sequences encoding the glutathione-S-transferase (GST) and an oligohistidine element, allowing expression of the corresponding fusion proteins; a phosp...

journal_title：BioTechniques

authors： Chatton B,Bahr A,Acker J,Kedinger C

更新日期：1995-01-01 00:00:00

abstract：:Haplotypes are useful in population genetics and medicine. Determining haplotypes in the absence of DNA samples from appropriate family members can be difficult and laborious. We have developed a rapid and reproducible method for haplotyping an individual in the absence of relatives. The method utilizes pairs of allel...

journal_title：BioTechniques

authors： Sarkar G,Sommer SS

更新日期：1991-04-01 00:00:00

abstract：:Few studies have focused on the significance of ras protein levels in human malignancy, in part because of the inherent difficulty in quantitation of the ras gene product. We have developed a method for the enzymatic determination of the ras gene product and have used this method for the quantitation of ras gene produ...

journal_title：BioTechniques

authors： Gutheil JC,Mane SM,Bass KA,Lee EJ,Needleman SW

更新日期：1990-08-01 00:00:00

abstract：:The ZEBRA protein is a transcriptional activator that induces expression of viral lytic genes in cells harboring latent Epstein-Barr virus (EBV). In this report it is shown that a derivative of ZEBRA that cannot activate transcription (Zd) can be used to detect and characterize activation domains. Three expression vec...

journal_title：BioTechniques

authors： Asković S,Baumann R

更新日期：1997-05-01 00:00:00

abstract：:Presepsin is a 13-kDa N-terminal glycoprotein of CD14. Previously, agitation-induced increases in presepsin levels have been reported; however, the mechanism remains poorly understood. In this study, we aimed to reveal the mechanism of presepsin increase. The agitated plasma or serum was separated using gel exclusion ...

journal_title：BioTechniques

authors： Morino G,Takahashi G,Kan S,Inoue Y,Sato K,Shirakawa K

更新日期：2021-01-29 00:00:00

abstract：:We describe the application of simple cloning by prolonged overlap extension for multiple site-directed mutagenesis in the same plasmid. We show that it is possible to use this technique with very short PCR templates. The technique is ideally suited for the generation of longer donor DNA sequences for CRISPR/Cas9-medi...

journal_title：BioTechniques

authors： Hejlesen R,Füchtbauer EM

更新日期：2020-06-01 00:00:00

abstract：:Microsatellite sequences are important markers for population genetics studies. In the past, the development of adequate microsatellite primers has been cumbersome. However with the advent of next-generation sequencing technologies, marker identification in genomes of non-model species has been greatly simplified. Her...

journal_title：BioTechniques

authors： Grohme MA,Soler RF,Wink M,Frohme M

更新日期：2013-11-01 00:00:00

abstract：:We report a simple and inexpensive method to quantitate loading and transfer of RNA and to examine RNA integrity for use with Northern hybridization. This technique involves quantitation by computer-assisted video densitometry of a negative photograph of 28S and 18S rRNA from ethidium bromide-stained RNA fixed to nylo...

journal_title：BioTechniques

authors： Correa-Rotter R,Mariash CN,Rosenberg ME

更新日期：1992-02-01 00:00:00

abstract：:Vascular targeting agents (VTAs) can be produced by linking antibodies or antibody fragments directed against endothelial cell markers to effector moieties. So far, it has been necessary to produce the components of VTAs (antibody, antibody fragment, linker, and effector) separately and, subsequently, to conjugate the...

journal_title：BioTechniques

authors： Gottstein C,Wels W,Ober B,Thorpe PE

更新日期：2001-01-01 00:00:00

abstract：:Loop-mediated isothermal amplification (LAMP) is a rapid and reliable sequence-specific isothermal nucleic acid amplification technique. To date, all reported real-time detection methods for LAMP have been restricted to single targets, limiting the utility of this technique. Here, we adapted standard LAMP primers to c...

journal_title：BioTechniques

authors： Tanner NA,Zhang Y,Evans TC Jr

更新日期：2012-08-01 00:00:00

abstract：:A novel method for large-scale plasmid preparation is described. Crude extracts are subjected to acidic phenol extraction to remove any contaminants present in the aqueous phase. The supercoiled plasmid DNA, which preferentially remains in the organic phase and inter-phase, is extracted back into the aqueous phase wit...

journal_title：BioTechniques

authors： Wang Z,Rossman TG

更新日期：1994-03-01 00:00:00

abstract：:A major problem in analyzing forensic casework samples is the presence of genetic material from more than one individual in the material evidence. For instance, in sexual assault cases the evidence (vaginal swabs) usually contains a majority of vaginal epithelial cells and varying amounts of sperm cells from the perpe...

journal_title：BioTechniques

authors： Allen M,Saldeen T,Gyllensten U

更新日期：1995-09-01 00:00:00

abstract：:A general method for solid-phase gene assembly on streptavidin-coated magnetic beads has been developed. The introduction of biotin in the 5'-end of the initiation oligonucleotide enables anchoring to the bead by means of the streptavidin-biotin interaction. The immobilization of one oligonucleotide enables controlled...

journal_title：BioTechniques

authors： Ståhl S,Hansson M,Ahlborg N,Nguyen TN,Liljeqvist S,Lundeberg J,Uhlén M

更新日期：1993-03-01 00:00:00

abstract：:An easy, sensitive and direct fluorescent immunodetection method for proteins is described using the new fluorochrome PBXL-1 imaged with the FMBIO II Laser Scanning Imaging System. PBXL-1 is derived from a protein supra-molecular complex that contains a large number of chromophores. This complex, the phycobilisome, is...

journal_title：BioTechniques

authors： Morseman JP,Moss MW,Zoha SJ,Allnutt FC

更新日期：1999-03-01 00:00:00

abstract：:SCORE, a program for computer-assisted scoring of Southern blots of clone DNA, retains the use of expert human judgment while taking over much of the drudgery of the scoring task. The primary functions of the program are to help make an aligned overlay of the fluorescence gel image and the autoradiogram blot image, to...

journal_title：BioTechniques

authors： Cannon TM,Koskela RJ,Burks C,Stallings RL,Ford AA,Hempfner PE,Brown HT,Fickett JW

更新日期：1991-06-01 00:00:00

abstract：:A technique that can be used to isolate vector/insert junctions from clones in vectors, such as yeast artificial chromosomes and P1s, and to sequence plasmid inserts more rapidly has been developed. A vector primer is combined with single, randomly chosen oligonucleotides in PCRs, to create pools of products. With 12-...

journal_title：BioTechniques

authors： Swensen J

更新日期：1996-03-01 00:00:00

abstract：:We have developed an automated purification method for dye-terminator-based DNA sequencing products using a magnetic bead approach. This 384-well protocol generates sequence fragments that are essentially free of template DNA, salt, and excess dye-terminator products. In comparison with traditional ethanol precipitati...

journal_title：BioTechniques

authors： Elkin C,Kapur H,Smith T,Humphries D,Pollard M,Hammon N,Hawkins T

更新日期：2002-06-01 00:00:00

abstract：:Quantitative PCR and reverse transcription PCR (RT-PCR) are widely used in biomedical, industrial and other research applications to determine the number of RNA or DNA molecules of a specific type and/or sequence in a sample of interest. We have developed an assay system to accurately quantitate PCR products that util...

journal_title：BioTechniques

authors： Martin CS,Butler L,Bronstein I

更新日期：1995-05-01 00:00:00

abstract：:Human embryonic kidney (HEK293) cells were stably transduced with a retroviral vector containing an expression cassette for a short-lived green fluorescent protein (d2EGFP) and the neomycin resistance gene (Neor). When Neor HEK293 clones were treated with proteasome inhibitors, lactacystin or MG132, an increase in the...

journal_title：BioTechniques

authors： Andreatta C,Nahreini P,Hovland AR,Kumar B,Edwards-Prasad J,Prasad KN

更新日期：2001-03-01 00:00:00

abstract：:Previous experiments have clearly demonstrated that microtubule dynamic instability is regulated in living cells, but the molecular mechanisms that are responsible for this regulation are not well understood. We describe two rapid, functional assays that can be used to screen cell extracts for regulators of microtubul...

journal_title：BioTechniques

authors： McMeekan J,Wadsworth P

更新日期：1998-05-01 00:00:00

abstract：:We systematically evaluated the performance and reliability of several widely used, commercially available actin-filament probes in a highly motile breast adenocarcinoma cell line to optimize the visualization of F-actin-rich dynamic lamellipodia. We evaluated four Phalloidin-fluorophores, two anti-actin antibodies, a...

journal_title：BioTechniques

authors： DesMarais V,Eddy RJ,Sharma VP,Stone O,Condeelis JS

更新日期：2019-03-01 00:00:00

abstract：:RNA interference (RNAi) is a recent advance that provides the possibility to reduce the expression of specific target genes in cultured mammalian cells with potential applications on a genome-wide scale. However, to achieve this, robust methodologies that allow automated and efficient delivery of small interfering RNA...

journal_title：BioTechniques

authors： Erfle H,Simpson JC,Bastiaens PI,Pepperkok R

更新日期：2004-09-01 00:00:00

abstract：:The extraction of high-quality nucleic acid may be problematic in formalin-fixed tissues because of cross-linking between proteins and DNA. Old fixed tissue specimens do produce fragmented DNA (<1.2 kb), which is only used for PCR amplification. Here we show that high molecular weight DNA (>194 kb) can be successfully...

journal_title：BioTechniques

authors： Fang SG,Wan QH,Fujihara N

更新日期：2002-09-01 00:00:00

abstract：:We have developed a novel method for the rapid preparation of large quantities of 3' end-labeled single-stranded (ss) DNA (ssDNA) probes for transcript mapping. A recombinant phagemid vector containing the probe sequence was used to raise large quantities of ssDNA. Based on the DNA sequence of the probe, an oligonucle...

journal_title：BioTechniques

authors： Silk GW,Wu M

更新日期：1993-05-01 00:00:00

abstract：:Under-agarose chemotaxis has been used previously to assess the ability of neutrophils to respond to gradients of chemoattractant. We have adapted this assay to the chemotactic movement of Dictyostelium amoebae in response to folic acid. Troughs are used instead of wells to increase the area along which the cells can ...

journal_title：BioTechniques

authors： Laevsky G,Knecht DA

更新日期：2001-11-01 00:00:00