Differential screening of a subtracted cDNA library: a method to search for genes preferentially expressed in multiple tissues.

abstract：:An improved method was developed for in situ reverse transcription-polymerase chain reaction (RT-PCR) to detect and localize mRNA in tissue sections. The coverslip mounted-immersion cycled (COSMIC) in situ RT-PCR technique combines the advantages of solution-phase PCR with the tissue immobilization necessary for in si...

journal_title：BioTechniques

authors： Ertsey R,Scavo LM

更新日期：1998-01-01 00:00:00

abstract：:We present a tool for dispensing very low volumes (20 nL or more) of ultra high viscosity (UHV) medical-grade alginate hydrogels. It uses a modified piezo-driven micrometering valve, integrated into a versatile system that allows fast prototyping of encapsulation procedures and scaffold production. Valves show excelle...

journal_title：BioTechniques

authors： Gepp MM,Ehrhart F,Shirley SG,Howitz S,Zimmermann H

更新日期：2009-01-01 00:00:00

abstract：:In the quest to move more basic research discoveries into the clinic, the medical community is building training programs to help researchers gain "translational" skills. Sarah Webb explores how PhDs are learning to speak the language of the clinic. ...

journal_title：BioTechniques

authors： Webb SA

更新日期：2014-08-01 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：:The oligonucleotide ligation assay (OLA) was adapted to the genotyping of the N-arylamine-acetyltransferase (NAT2) gene. This assay allows the use of 96-well microplates and robotic workstations for high sample throughput. We found this assay to be accurate, efficient and reliable. Another advantage for epidemiologica...

journal_title：BioTechniques

authors： Bigler J,Chen C,Potter JD

更新日期：1997-04-01 00:00:00

abstract：:Inverse PCR has been used for the recovery of genome regions flanking a known sequence, although its application to metagenome walking is limited due to inefficient amplification from low copy number fragments. Here we present an improved inverse PCR scheme that enables walking of rare fragments in environmental metag...

journal_title：BioTechniques

authors： Uchiyama T,Watanabe K

更新日期：2006-08-01 00:00:00

abstract：:The need for a primer hybridization step before sequencing has been eliminated using a stem-loop structure generated by PCR. The loop structure is obtained by careful design of the PCR primer or by cloning the target DNA into a dedicated vector (pRIT 28HP). After solid-phase capture of the PCR product, the loop is for...

journal_title：BioTechniques

authors： Ronaghi M,Pettersson B,Uhlén M,Nyrén P

更新日期：1998-11-01 00:00:00

abstract：:Sample automation and management is increasingly important as the number and size of population-scale and high-throughput projects grow. This is particularly the case in large-scale population studies where sample size is far outpacing the commonly used 96-well plate format. To facilitate management and transfer of sa...

journal_title：BioTechniques

authors： Cario CL,Witte JS

更新日期：2018-12-01 00:00:00

abstract：:Comprehensive analysis of DNA methylation patterns is critical for understanding the molecular basis of many human diseases. While hundreds of PCR-based DNA methylation studies are published every year, the selection and implementation of appropriate methods for these studies can be challenging for molecular genetics ...

journal_title：BioTechniques

authors： Hernández HG,Tse MY,Pang SC,Arboleda H,Forero DA

更新日期：2013-10-01 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：: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：:Real-time reverse transcription PCR (RT-PCR) is a sensitive and accurate method to monitor gene expression and is often used to profile the expression of putative tumor antigens in the context of immunotherapy. However, this technique consists of several steps, including cell processing, RNA extraction, RNA storage, a...

journal_title：BioTechniques

authors： Aerts JL,Gonzales MI,Topalian SL

更新日期：2004-01-01 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 cost-effective method for the synthesis of an internal fluorescently labeled DNA standard for fragment sizing using an automated DNA sequencer. A set of primer pairs labeled with ROX was developed to amplify 12 DNA fragments, 58-417 bp, derived from a conserved region of plant chloroplast DNA....

journal_title：BioTechniques

authors： Brondani RP,Grattapaglia D

更新日期：2001-10-01 00:00:00

abstract：:Selective delivery of genes to ocular tissues in vivo has been a long sought after goal for potential gene therapy of ocular disease. The gene gun was considered for this purpose because of its ability to focally transfer DNA to cells through gold microparticles coated with DNA. Through experimentation, we optimized a...

journal_title：BioTechniques

authors： Tanelian DL,Barry MA,Johnston SA,Le T,Smith G

更新日期：1997-09-01 00:00:00

abstract：:Ribosome profiling is a powerful tool for characterizing in vivo protein translation at the genome scale, with multiple applications ranging from detailed molecular mechanisms to systems-level predictive modeling. Though highly effective, this intricate technique has yet to become widely used in the microbial research...

journal_title：BioTechniques

authors： Latif H,Szubin R,Tan J,Brunk E,Lechner A,Zengler K,Palsson BO

更新日期：2015-06-01 00:00:00

abstract：:Characterization of the thermodynamics of binding interactions is important in improving our understanding of bimolecular recognition and forms an essential part of the rational drug design process. Isothermal titration calorimetry (ITC) is rapidly becoming established as the method of choice for undertaking such stud...

journal_title：BioTechniques

authors： Holdgate GA

更新日期：2001-07-01 00:00:00

abstract：:A simple spectrophotometric method for measuring DNA in proliferating cells is described. The method is an adaptation of the widely used diphenylamine (DPA) reaction to examine DNA in cells grown in a 96-well plate. This assay was capable of detecting as little as 10 ng DNA and could be used to measure DNA in stored a...

journal_title：BioTechniques

authors： Natarajan N,Shambaugh GE 3rd,Elseth KM,Haines GK,Radosevich JA

更新日期：1994-07-01 00:00:00

abstract：:The type IIS/IIC restriction endonuclease TspGWI recognizes the sequence 5'-ACGGA-3', cleaving DNA 11/9 nucleotides downstream. Here we show that sinefungin, a cofactor analog of S-adenosyl methionine, induces a unique type of relaxation in DNA recognition specificity. In the presence of sinefungin, TspGWI recognizes ...

journal_title：BioTechniques

authors： Zylicz-Stachula A,Zołnierkiewicz O,Jeżewska-Frąckowiak J,Skowron PM

更新日期：2011-06-01 00:00:00

abstract：:The technique described here is a fast and simple method of extracting chloroplast DNA (cpDNA). It overcomes the need for differential centrifugation using density gradients. The leaves do not have to be kept in the dark and lyophilized before extraction, but lyophilization is still possible. The chloroplasts are spec...

journal_title：BioTechniques

authors： Mariac C,Trouslot P,Poteaux C,Bezançon G,Renno JF

更新日期：2000-01-01 00:00:00

abstract：:Much effort has been directed toward the isolation and characterization of homeobox cDNAs from numerous cell types because they encode transcription factors important to many cellular processes, including pattern formation in the embryo, cell growth and cell differentiation. Many novel homeobox cDNAs have been isolate...

journal_title：BioTechniques

authors： Gorski DH,LePage DF,Walsh K

更新日期：1994-05-01 00:00:00

abstract：:A complete image digitizing and processing system is described for capturing, enhancing and analyzing molecular fingerprints. The low-cost, high-resolution system features a Motorola 68000 processor, multi-tasking, a separate video coprocessor, and color or gray scale processing. Thousands of manipulations are possibl...

journal_title：BioTechniques

authors： Mitchell BW,Palmieri S

更新日期：1990-10-01 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：:A pipeline has been created for the characterization of Arabidopsis thaliana mutants by generating flanking sequence tags (FSTs) and optimized for economic, high-throughput production. The GABI-Kat collection of T-DNA mutagenized A. thaliana plants was used as a source of independent transgenic lines. The pipeline inc...

journal_title：BioTechniques

authors： Strizhov N,Li Y,Rosso MG,Viehoever P,Dekker KA,Weisshaar B

更新日期：2003-12-01 00:00:00

abstract：:A method is described for preparing mutants with multiple, site-directed mutations by ordered coupling of PCR-generated fragments catalyzed by a thermostable DNA ligase. Annealing of the sense strands of the fragments to a single-stranded (antisense) template created a full-length sense strand leaving only nicks that ...

journal_title：BioTechniques

authors： Rouwendal GJ,Wolbert EJ,Zwiers LH,Springer J

更新日期：1993-07-01 00:00:00

abstract：:beta-galactosidase (beta-gal), the product of the E. coli LacZ gene, has been used extensively as a reporter in numerous systems. Until recently, the most commonly used method of detecting beta-gal reporter enzymatic activity was a colormetric assay based on the cleavage of the beta-gal substrate 5-bromo-4-chloro-3-in...

journal_title：BioTechniques

authors： Nazarenko DA,Dertinger SD,Gasiewicz TA

更新日期：2001-04-01 00:00:00

abstract：:Cell-based immunizations are often used when membrane antigens are difficult to purify. To confirm that an antibody binding to the surface of a cell line is, in fact, binding to the desired antigen, FACS can be performed independently on two cell lines, a transfected cell line expressing the antigen of interest and a ...

journal_title：BioTechniques

authors： Yang XP,Gallo M,Ngan I,Nocerini M,Chen MM

更新日期：2002-03-01 00:00:00

abstract：:We have developed an improved subtractive hybridization method that provides a fast, simple and reliable isolation of desired different sequences from two compared DNA libraries, one of which contains all unwanted homologues (subtracter) and another contains certain desired heterologues (tester). The DNA library can b...

journal_title：BioTechniques

authors： Ying SY,Lin S

更新日期：1999-05-01 00:00:00

abstract：:In the design of oligonucleotide sequences for targeting DNA or RNA sequences, it can be difficult to identify sequences that will hybridize only to the intended target. The term "sequence-specific" or "sequence-nonspecific" is often used to describe the interactions of an oligonucleotide with a mixture of DNA or RNA....

journal_title：BioTechniques

authors： Hyndman D,Cooper A,Pruzinsky S,Coad D,Mitsuhashi M

更新日期：1996-06-01 00:00:00

abstract：:Here we present a procedure for preparing amino acid mixtures--having both the desired composition and a physiological pH--at high concentrations for cell-free expression systems. Up to 2.1 mg/mL of active protein was synthesized in batch mode reactions with an all Escherichia coli cell-free expression system. Our met...

journal_title：BioTechniques

authors： Caschera F,Noireaux V

更新日期：2015-01-01 00:00:00