A simple procedure for recovering the denaturing effect of methylmercury in agarose gel electrophoresis.

abstract：:14CO2 capture is commonly used to evaluate the cellular oxidation rate of respiratory substrates. A modification of the established 14CO2-capture method was developed that enables the use of cells in adherent culture and easy analysis of multiple samples under different culture conditions. The use of commercially avai...

journal_title：BioTechniques

authors： Collins CL,Bode BP,Souba WW,Abcouwer SF

更新日期：1998-05-01 00:00:00

abstract：:Plasmid pUC18rspL is a 3.788-kilobase pair vector, derived from pUC18, pKK232-8 and pHSG664, which identifies promoter-bearing DNA fragments functional in StrA E. coli by activation of a promoterless streptomycin-sensitive gene cartridge (rspL). Expression of the plasmid-borne rspL gene leads to sensitivity dominance ...

journal_title：BioTechniques

authors： Vockley JG,Pène JJ

更新日期：1990-12-01 00:00:00

abstract：:Pairs of primers flanking known miniTn10 transposon insertion sites were used to confirm the presence of the transposon in DNA isolated from Legionella pneumophila mutants. It was expected that the polymerase chain reaction products derived from the mutant template would be larger than those from the wild-type (WT) te...

journal_title：BioTechniques

authors： Viswanathan VK,Krcmarik K,Cianciotto NP

更新日期：1999-09-01 00:00:00

abstract：:Thymidine kinase 1 (TK1) is an enzyme involved in DNA precursor synthesis that has been used as a biomarker for prognosis and monitoring of different malignancies. In this study, we compared two immunoassays for measuring TK1 protein concentrations: the TK 210 ELISA (AroCell AB) and TK1 ELISA from Abcam. Overall, the ...

journal_title：BioTechniques

authors： Kumar JK,Holmgren S,Levedahl KH,Höglund M,Venge P,Eriksson S

更新日期：2020-06-01 00:00:00

abstract：:Currently, in real-time PCR, one often has to choose between using a sequence-specific probe and a nonspecific double-stranded DNA (dsDNA) binding dye for the detection of amplified DNA products. The sequence-specific probe has the advantage that it only detects the targeted product, while the nonspecific dye has the ...

journal_title：BioTechniques

authors： Lind K,Ståhlberg A,Zoric N,Kubista M

更新日期：2006-03-01 00:00:00

abstract：:We describe a simple and efficient system for epitope mapping by cloning random gene fragments into a specially designed gIIIp-based phage display vector. DNA encoding the antigen of interest is PCR-amplified and partially digested with DNaseI to generate 50-150-bp-long fragments, which are polished with T4 DNA polyme...

journal_title：BioTechniques

authors： Gupta S,Arora K,Sampath A,Khurana S,Singh SS,Gupta A,Chaudhary VK

更新日期：1999-08-01 00:00:00

abstract：:In this study, we have developed an air-interface culture system in which guinea pig tracheobronchial epithelial (GPTE) cells rapidly form tight monolayers. Enzymatically isolated GPTE cells were plated on collagen-treated polycarbonate microporous cell culture inserts at a density of 10(6) cells/cm2 (day 0). Bioelect...

journal_title：BioTechniques

authors： Robison TW,Dorio RJ,Kim KJ

更新日期：1993-09-01 00:00:00

abstract：:Here we introduce a modified antibody staining method that uses up to 80% less antibody for flow cytometry. We demonstrate this method for the detection of antigens expressed at high, moderate, or low levels in mouse and rat lymphocytes as well as mouse mammary epithelial cells. We obtained reproducibly accurate resul...

journal_title：BioTechniques

authors： Sharma D,Eichelberg MR,Haag JD,Meilahn AL,Muelbl MJ,Schell K,Smits BM,Gould MN

更新日期：2012-07-01 00:00:00

abstract：:The PCR method has proved to be an invaluable tool for the specific and sensitive detection of genetically modified material (e.g., Roundup Ready Soybean and Bt-176 "Maximizer" Maize) in foodstuffs. The first step in the procedure, namely the purification of nucleic acids from the sample, is often the deciding factor ...

journal_title：BioTechniques

authors： Tengel C,Schüssler P,Setzke E,Balles J,Sprenger-Haussels M

更新日期：2001-08-01 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：: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：:The concentration of proteins in cells is an important parameter that determines how a protein will interact with other proteins or pharmacological agents. Recent developments in Western blotting techniques have now made this a method of choice to measure protein concentration in complex solutions such as total cell e...

journal_title：BioTechniques

authors： Fradelizi J,Friederich E,Beckerle MC,Golsteyn RM

更新日期：1999-03-01 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：:Chitin from crustacean shells has often been used to isolate and purify plant lectins that have an affinity for poly-N-acetylglucosamine (poly-GlcNAc). When we used washed chitin from crab shells as an affinity medium to isolate a lectin from Pinus strobus L. (eastern white pine) ovules, we found that a substance havi...

journal_title：BioTechniques

authors： Whitmore FA

更新日期：1992-02-01 00:00:00

abstract：:Here we describe a method for the isolation of PCR-ready genomic DNA from various zebrafish tissues that is based on a previously published murine protocol. The DNA solutions are of sufficient quality to allow PCR detection of transgenes from all commonly used zebrafish tissues. In sperm, transgene amplification was s...

journal_title：BioTechniques

authors： Meeker ND,Hutchinson SA,Ho L,Trede NS

更新日期：2007-11-01 00:00:00

abstract：:The Drosophila heart has gained considerable traction as a model of cardiac development and physiology. Previously we described a semiautomatic optical heartbeat analysis (SOHA) method for quantifying functional parameters from the fly heart that facilitated its use as an organ system and disease model. Here we presen...

journal_title：BioTechniques

authors： Cammarato A,Ocorr S,Ocorr K

更新日期：2015-02-01 00:00:00

abstract：:Replication studies on human immunodeficiency virus 1 (HIV-1) rely on a few laboratory strains that are divergent from dominant HIV-1 subtypes in the epidemic. Several phenotypic differences between diverse HIV-1 isolates and subtypes could affect vaccine development and treatment, but this research field lacks robust...

journal_title：BioTechniques

authors： Dudley DM,Gao Y,Nelson KN,Henry KR,Nankya I,Gibson RM,Arts EJ

更新日期：2009-05-01 00:00:00

abstract：:Microarrays printed on glass slides are often constructed by covalently linking modified oligonucleotide probes to a derivatized surface at considerable expense. In this article, we demonstrate that 14-base oligonucleotides with a poly(T)10 - poly(C)10 tail (TC tag), but otherwise unmodified, can be linked by UV light...

journal_title：BioTechniques

authors： Gudnason H,Dufva M,Duong Bang D,Wolff A

更新日期：2008-09-01 00:00:00

abstract：:Complete brain fixation can be achieved with transthoracic cardiac infusion without thoracotomy. Light and electron microscopy tissue sections reveal preservation of cytoplasmic and nuclear structure at all magnification levels. Punched samples were obtained from the fixed tissue specimens in precisely localized areas...

journal_title：BioTechniques

authors： Eichenbaum KD,Eichenbaum JW,Fadiel A,Miller DC,Demir N,Naftolin F,Stern A,Pevsner PH

更新日期：2005-10-01 00:00:00

abstract：:GC-rich DNA regions were PCR-amplified with Taq DNA polymerase using either the canonical set of deoxynucleoside triphosphates or mixtures in which the dCTP had been partially or completely replaced by its N4-methylated analog, N4-methyl-2'-deoxycytidine 5'-triphosphate (N4me-dCTP). In the case of a particularly GC-ri...

journal_title：BioTechniques

authors： Flores-Juárez CR,González-Jasso E,Antaramian A,Pless RC

更新日期：2016-10-01 00:00:00

abstract：:A direct method to study essential genes is to construct conditional knock-down mutants by replacement of their native promoter by an inducible one. In Mycobacteria, replacement of an essential gene promoter with an anhydrotetracycline inducible one was successfully used but required a multi-step approach. In this wor...

journal_title：BioTechniques

authors： Texier P,Coddeville M,Bordes P,Genevaux P

更新日期：2018-09-01 00:00:00

abstract：:Genome variation provides researchers with thousands of markers with which to study human demographic history and phenotypes. Insertion-deletion (indel) polymorphism is an important and abundant form of human genome variation, and convenient methods for genotyping indels are therefore needed. Here we evaluate dynamic ...

journal_title：BioTechniques

authors： Sawyer SL,Howell WM,Brookes AJ

更新日期：2003-08-01 00:00:00

abstract：:Here we describe a convenient method to generate homologous recombinant baculoviral genomes in E. coli. The recombination takes place with the aid of recombination enzymes provided by the phage lambda Red system between a bacmid (a baculoviral genome that can replicate in bacteria) and a linear fragment. Proof of conc...

journal_title：BioTechniques

authors： Hou S,Chen X,Wang H,Tao M,Hu Z

更新日期：2002-04-01 00:00:00

abstract：:Subtractive hybridization has been widely used for the identification of differentially expressed genes. Here we describe a simple, sensitive strategy of subtractive hybridization that involves binding the driver poly(A)+ RNA pool to paramagnetic Dynabeads Oligo (dT)25. After hybridization with target cDNA, the molecu...

journal_title：BioTechniques

authors： Mészáros M,Morton DB

更新日期：1996-03-01 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：:Conformation-sensitive gel electrophoresis (CSGE) has been introduced as the most reliable method for the screening of large and multi-exon genes because of its simplicity, sensitivity and specificity. Based on heteroduplex formation and with the use of mildly denaturing solvents, it allows detection of single-base mu...

journal_title：BioTechniques

authors： Blesa JR,Hernández-Yago J

更新日期：2000-05-01 00:00:00

abstract：:We have compared RNA polymerase promoter activities in PCR-generated DNA fragments for use in the in vitro transcription of cRNA probes. Sense oligonucleotide primers, specific for the mouse acidic fibroblast growth factor gene, were synthesized with 5' extensions containing promoter sequences for the T7, T3 and SP6 R...

journal_title：BioTechniques

authors： Logel J,Dill D,Leonard S

更新日期：1992-10-01 00:00:00

abstract：:Several DNA extraction methods have been reported for use with digesta or fecal samples, but problems are often encountered in terms of relatively low DNA yields and/or recovering DNA free of inhibitory substances. Here we report a modified method to extract PCR-quality microbial community DNA from these types of samp...

journal_title：BioTechniques

authors： Yu Z,Morrison M

更新日期：2004-05-01 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：:A fully automated nucleic acid analysis system is described, which offers positive sample identification, improved sensitivity and reduced user interaction compared to conventional techniques. The system relies on the sequence-specific capture of DNA onto solid-phase particles, confirming product identity without the ...

journal_title：BioTechniques

authors： Brown A,Akinsanya AA,Barker SJ,Brophy M,Dobb AK,Doyle SM,Hudson IR,Minter SJ,Wraith MJ,Oultram JD

更新日期：1999-07-01 00:00:00