Amplification of mRNA populations by a cDNA tag strategy.

abstract：:We have developed a rapid [3H]colchicine competition-binding scintillation proximity assay (SPA) to evaluate antimitotic compounds that bind to the colchicine-binding site on tubulin. The premise of our assay is that compounds will compete with radiolabeled colchicine for the tubulin-binding domain. Biotin-labeled tub...

journal_title：BioTechniques

authors： Tahir SK,Kovar P,Rosenberg SH,Ng SC

更新日期：2000-07-01 00:00:00

abstract：:Advances in in vitro display and protein engineering yield therapeutics with affinities in the picomolar range. The Gyrolab® microfluidics platform uses the kinetic exclusion assay principle to measure subnanomolar solution affinities. This work describes application of the Gyrolab solution affinity module and the new...

journal_title：BioTechniques

authors： Dai Z,Juneja J,Schneeweis L,Cohen D,Marsilio F,Morin P,DasGupta R

更新日期：2020-09-01 00:00:00

abstract：:PhDs, postdocs, fellows and newly appointed lecturers all fall into the category of early-career researchers (ECRs). The life of researchers in early stages of their career is far from straightforward. Funding is a huge barrier for many ECRs looking to make the next step in their career, but it's not the only challeng...

journal_title：BioTechniques

authors：

更新日期：2019-01-01 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：:Genetic immunization is an attractive approach to generate antibodies because native proteins are expressed in vivo with normal posttranscriptional modifications, avoiding time-consuming and costly antigen isolation or synthesis. Hydrodynamic tail or limb vein delivery of naked plasmid DNA expression vectors was used ...

journal_title：BioTechniques

authors： Bates MK,Zhang G,Sebestyén MG,Neal ZC,Wolff JA,Herweijer H

更新日期：2006-02-01 00:00:00

abstract：:Xer-cise is a technique using antibiotic resistance cassettes flanked by dif sites allowing spontaneous and accurate excision from bacterial chromosomes with a high frequency through the action of the cellular recombinase XerCD. Here, we report a significant improvement of Xer-cise in Mycobacteria. Zeocin resistance c...

journal_title：BioTechniques

authors： Boudehen YM,Wallat M,Rousseau P,Neyrolles O,Gutierrez C

更新日期：2020-02-01 00:00:00

abstract：:Here, we present a rapid and reproducible procedure based on square-wave pulse electroporation that allows efficient penetration of synthetic oligonucleotides into intact yeast cells. This procedure was successfully used to modify the yeast genome with small amounts of oligonucleotide. ...

journal_title：BioTechniques

authors： Barre FX,Mir LM,Lécluse Y,Harel-Bellan A

更新日期：1998-08-01 00:00:00

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：:The transcription factor GATA-1 is a zinc finger DNA-binding protein essential for the development of red blood cells. When we expressed different regions of the zinc finger domain in bacteria using an isopropyl-beta-D-thiogalactoside (IPTG) inducible system, growth of bacteria harboring the active DNA-binding domain ...

journal_title：BioTechniques

authors： Trudel P,Provost S,Massie B,Chartrand P,Wall L

更新日期：1996-04-01 00:00:00

abstract：:An assay is described in which an oligonucleotide probe is specifically digested by lambda exonuclease only when it is annealed to its complementary sequence. In this assay, a cycling effect occurs whereby a small amount of target sequence acts as a specific co-factor in the enzymatic degradation of a larger number of...

journal_title：BioTechniques

authors： Copley CG,Boot C

更新日期：1992-12-01 00:00:00

abstract：:A simple, non-destructive procedure is described to determine the quality of DNA arrays before they are used. It consists of a preliminary staining step of the DNA microarray by using SYBR green II, a fluorophore with specific affinity for ssDNA, followed by a laser scan analysis. The surface quality, integrity and ho...

journal_title：BioTechniques

authors： Battaglia C,Salani G,Consolandi C,Bernardi LR,De Bellis G

更新日期：2000-07-01 00:00:00

abstract：:DNA sequences containing tandem dinucleotide repeats represent an abundant source of DNA polymorphism in human and other eukaryotic genomes. Here we describe a novel technique for the identification and characterization of regions of DNA containing these repetitive elements. Using primers designed to recognize tandem ...

journal_title：BioTechniques

authors： Grist SA,Firgaira FA,Morley AA

更新日期：1993-08-01 00:00:00

abstract：:A new qualitative PCR product detection assay called competitive amplified single mutation detection by selective probe hybridization immunoassay (CASSI) was developed for genotyping the most common apolipoprotein E (apoE) polymorphisms. Single target DNA strands immobilized using biotin on streptavidin-coated micropl...

journal_title：BioTechniques

authors： Köhler T,Rost AK,Purschwitz K,Vondran S,Remke H,Wagner O,Richter V

更新日期：1998-07-01 00:00:00

abstract：:Current protocols for screening proliferative factors for β cells ex vivo are time-consuming, require cell lines or dissociated islets, and often entail expensive specialized screening equipment. Here we present an efficient and lower cost alternative that utilizes intact mouse islets for the initial screening of prol...

journal_title：BioTechniques

authors： Mosser RE,Gannon M

更新日期：2013-12-01 00:00:00

abstract：:A low order neural network-based filter was designed as a rapid screening agent for single-spanning transmembrane regions in an integrated informatics system. A rapid screening algorithm was seen as a compromise between costly structure-specific techniques and simple rules that gave a high false-positive rate for cDNA...

journal_title：BioTechniques

authors： Huang GM,Farkas J,Hood L

更新日期：1996-12-01 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：:We have developed a new strategy for differential screening of genes that are expressed in two or more tissues, and have used it to identify genes that are preferentially expressed in both testis and ovary. In this approach, testis-specific cDNAs were first generated using the suppression subtractive hybridization tec...

journal_title：BioTechniques

authors： Jin H,Cheng X,Diatchenko L,Siebert PD,Huang CC

更新日期：1997-12-01 00:00:00

abstract：:Integral membrane G protein-coupled receptors (GPCRs) compose the single most prolific class of drug targets, yet significant functional and structural questions remain unanswered for this superfamily. A primary reason for this gap in understanding arises from the difficulty of forming soluble, monodisperse receptor m...

journal_title：BioTechniques

authors： Leitz AJ,Bayburt TH,Barnakov AN,Springer BA,Sligar SG

更新日期：2006-05-01 00:00:00

abstract：:For a feasible and cost-effective impedance measurement of cellular alterations in real-time, we combined commercially available microelectrode arrays (MEAs), consisting of 60 microelectrodes, with a conventional impedance analyzer. For proof of principle, a breast carcinoma cell line (MCF-7) was cultured on MEAs, and...

journal_title：BioTechniques

authors： Rothermel A,Nieber M,Müller J,Wolf P,Schmidt M,Robitzki AA

更新日期：2006-10-01 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：:Cysts of free-living protozoa have an impact on the ecology and epidemiology of bacteria because they may act as a transmission vector or shelter the bacteria against hash environmental conditions. Detection and localization of intracystic bacteria and examination of the en- and excystment dynamics is a major challeng...

journal_title：BioTechniques

authors： Lambrecht E,Baré J,Claeys M,Chavatte N,Bert W,Sabbe K,Houf K

更新日期：2015-04-01 00:00:00

abstract：:Rapid identification of viruses is needed to monitor the blood supply for emerging threats. Here we present a method that meets these criteria and allows for the shotgun sequencing of novel, uncultured DNA viruses directly from human blood. This method employs selection based on the physical properties of viruses comb...

journal_title：BioTechniques

authors： Breitbart M,Rohwer F

更新日期：2005-11-01 00:00:00

abstract：:A method for cloning single-stranded oligonucleotides in a plasmid vector has been developed. The method relies on ligation of the oligonucleotide into suitable restriction enzyme sites of the cloning vector such that the site at the 5' end has a 5' overhang [for example, a Bgl II site (A decreases GATCT)], and the si...

journal_title：BioTechniques

authors： Mounts P,Wu TC,Peden K

更新日期：1989-04-01 00:00:00

abstract：:Francisella tularensis is one of the most deadly bacterial agents, yet most of the genetic determinants of pathogenesis are still unknown. We have developed an efficient targeted mutagenesis strategy in the model organism F. tularensis subsp. novicida by utilizing universal priming of optimized antibiotic resistance c...

journal_title：BioTechniques

authors： Liu J,Zogaj X,Barker JR,Klose KE

更新日期：2007-10-01 00:00:00

abstract：:A brief description of the setup of an inexpensive densitometric analysis system is provided. This system uses a software package and scanner that can be obtained for a total of less than $1000 and that can be operated by any Macintosh computer, including a MacPlus. In the event that a scanner is already available, th...

journal_title：BioTechniques

authors： Shea TB

更新日期：1994-06-01 00:00:00

abstract：:A method for conserving primers and differently labeled fluorogenic hydrolysis (i.e., TaqMan) probes at ambient conditions is presented. Primers and hydrolysis probes with four different fluorophore-quencher combinations (6- FAM-BHQ1, HEX-BHQ1, ROX-BHQ650, and Cy5-BHQ2) were mixed with trehalose and xanthan at final c...

journal_title：BioTechniques

authors： Rombach M,Kosse D,Faltin B,Wadle S,Roth G,Zengerle R,von Stetten F

更新日期：2014-09-01 00:00:00

abstract：:We describe a simple and efficient technique that facilitates the amplification of specific mRNA for cloning and sequencing purposes. An mRNA bound to a small piece of membrane filter is used as a template to synthesize complementary DNA. The product of this reaction is then transferred to a new tube and amplified usi...

journal_title：BioTechniques

authors： Ruiz A,Bok D

更新日期：1993-11-01 00:00:00

abstract：:Methods have been developed to rapidly visualize the size distribution of third complementarity-determining regions (CDR3) in immunoglobulin (Ig) and T-cell receptor (TCR) molecules. DNA fragments spanning the Ig or TCR CDR3 are generated by PCR using primers at fixed positions in the variable and constant segments. T...

journal_title：BioTechniques

authors： Raaphorst FM,Tami J,Sanz IE

更新日期：1996-01-01 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：:Previous work from this laboratory has shown that immunoporation has the potential for the selective transfection of a range of different animal cells based on their immunological identity. The unique ability of immunoporation to target cells for transfection combined with the high efficiency of transfection and the h...

journal_title：BioTechniques

authors： Tzavelas C,Bildirici L,Rickwood D

更新日期：2004-08-01 00:00:00