A protocol for the purification of bovine serum albumin free of deoxyribonuclease activity.

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：: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：:Reporter assays that use luciferase are widely employed for monitoring cellular events associated with gene expression. In general, firefly luciferase and Renilla luciferase are used for monitoring single gene expression. However, the expression of more than one gene cannot be monitored simultaneously by this system b...

journal_title：BioTechniques

authors： Nakajima Y,Kimura T,Sugata K,Enomoto T,Asakawa A,Kubota H,Ikeda M,Ohmiya Y

更新日期：2005-06-01 00:00:00

abstract：:To move microarray technology into the diagnostic realm, the impact of technical parameters, such as sample preparation and RNA extraction, needs to be understood and minimized. We evaluated the impact of two RNA extraction methods, DNase treatment and the amount of hybridized cDNA probe, on the outcome of microarray ...

journal_title：BioTechniques

authors： Ojaniemi H,Evengard B,Lee DR,Unger ER,Vernon SD

更新日期：2003-11-01 00:00:00

abstract：:In this study, we developed a method that allows cDNA library construction from a small amount of RNA without causing serious size bias in the resulting cDNA population. For this purpose, we adopted two-round cRNA amplification by T7 and SP6 RNA polymerases. The first-round cDNAs, flanked by the promoter sequences of ...

journal_title：BioTechniques

authors： Ohara R,Kikuno RF,Kitamura H,Ohara O

更新日期：2005-03-01 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：:Site-specific mutagenesis and directional subcloning were accomplished by using the polymerase chain reaction to generate products that can recombine to form circular DNA. This DNA was transfected into E. coli without phosphorylation of primers, restriction enzyme digestion or ligation. Specifically, the polymerase ch...

journal_title：BioTechniques

authors： Jones DH,Howard BH

更新日期：1990-02-01 00:00:00

abstract：:Normalization is a critical step in the analysis of microarray gene expression data. For dual-labeled array, traditional normalization methods assume that the majority of genes are non-differentially expressed and that the number of overexpressed genes approximately equals the number of under-expressed genes. However,...

journal_title：BioTechniques

authors： Wu Y,Yan L,Liu H,Sun H,Xie H

更新日期：2009-08-01 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：:Genetic immunization is a simple method for producing polyclonal antibodies in mice. To test if this approach could be used for monoclonal antibody production, biolistic transfection was used to immunize a mouse. High levels of polyclonal antibodies against human growth hormone (hGH) were elicited following three inoc...

journal_title：BioTechniques

authors： Barry MA,Barry ME,Johnston SA

更新日期：1994-04-01 00:00:00

abstract：:Caspase-3 is a key effector caspase that is activated in both extrinsic and intrinsic pathways of apoptosis. Available fluorescent sensors for caspase-3 activity operate in relatively short wavelength regions and are nonoptimal for multiparameter microscopy and whole-body imaging. In the present work, we developed new...

journal_title：BioTechniques

authors： Zlobovskaya OA,Sergeeva TF,Shirmanova MV,Dudenkova VV,Sharonov GV,Zagaynova EV,Lukyanov KA

更新日期：2016-02-01 00:00:00

abstract：:Intrinsically disordered proteins (IDPs) are subject to post-translational modifications. This allows the same polypeptide to be involved in different interaction networks with different consequences, ranging from regulatory signalling networks to the formation of membrane-less organelles. We report a robust method fo...

journal_title：BioTechniques

authors： Brázda P,Šedo O,Kubíček K,Štefl R

更新日期：2019-07-01 00:00:00

abstract：:The degree of fluorescence polarization (FP) of a fluorescent molecule is a reflection of its molecular weight (Mr). FP is therefore a useful detection methodfor homogeneous assays in which the starting reagents and products differ significantly in Mr. We have previously shown that FP is a good detection method for th...

journal_title：BioTechniques

authors： Hsu TM,Chen X,Duan S,Miller RD,Kwok PY

更新日期：2001-09-01 00:00:00

abstract：:Here we report a simple new method for exposing cells to normoxic and hypoxic conditions using vacuum bags, normally employed for food storage, to establish and maintain low oxygen levels in vitro. Vacuum bags were gassed with a mixture containing specified levels of oxygen, then sealed, creating a hypoxic microenviro...

journal_title：BioTechniques

authors： Bakmiwewa SM,Heng B,Guillemin GJ,Ball HJ,Hunt NH

更新日期：2015-10-01 00:00:00

abstract：:A simple and rapid procedure for recovering the denaturing effect of methylmercuric hydroxide in agarose gel electrophoresis is described. The procedure consisted of the treatment of the commercial methylmercuric hydroxide solutions with Amberlite, a mixture of anion- and cation-exchange resins. This treatment greatly...

journal_title：BioTechniques

authors： Fernández-Silva P,Enriquez JA,Montoya J

更新日期：1992-04-01 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：:Random mutagenesis followed by an in vitro selection procedure was shown to be capable of identifying important bases of the hairpin ribozyme for cleavage of an RNA target sequence. The selection scheme enriched the RNA population for those molecules capable of efficient site-specific self-cleavage in the absence of l...

journal_title：BioTechniques

authors： Siwkowski A,Humphrey M,De-Young MB,Hampel A

更新日期：1998-02-01 00:00:00

abstract：:Thin spun-coat films (~4 nm thick) of graphene oxide (GO) constitute a versatile surface chemistry compatible with a broad range of technologically important sensor materials. Countless publications are dedicated to the nuances of surface chemistries that have been developed for sensors, with almost every material hav...

journal_title：BioTechniques

authors： Mulvaney SP,Stine R,Long NC,Tamanaha CR,Sheehan PE

更新日期：2014-07-01 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：:One of the challenges of performing live-cell imaging in plants is establishing a system for securing the sample during imaging that allows for the rapid addition of treatments. Here we report how a commercially available device called a HybriWell™ can be repurposed to create an imaging chamber suitable for Arabidopsi...

journal_title：BioTechniques

authors： Vang S,Seitz K,Krysan PJ

更新日期：2018-06-01 00:00:00

abstract：:High-throughput genotyping technologies such as DNA pooling and DNA microarrays mean that whole-genome screens are now practical for complex disease gene discovery using association studies. Because it is currently impractical to use all available markers, a subset is typically selected on the basis of required satura...

journal_title：BioTechniques

authors： Simpson CL,Hansen VK,Sham PC,Collins A,Powell JF,Al-Chalabi A

更新日期：2004-12-01 00:00:00

abstract：:We have developed a high-throughput, multiplex reverse transcription PCR (RTPCR) assay that is suitable for the analysis of medium-to low-copy cellular RNA transcripts from small numbers of cells (10(4)). High throughput was attained by utilizing microplate-based RNA extraction and RTPCR protocols, followed by PCR pro...

journal_title：BioTechniques

authors： Su S,Vivier RG,Dickson MC,Thomas N,Kendrick MK,Williamson NM,Anson JG,Houston JG,Craig FF

更新日期：1997-06-01 00:00:00

abstract：:The expression of foreign proteins in Saccharomyces cerevisiae is a powerful tool for basic research and the biotechnological industry. In spite of the potential of S. cerevisiae, only a few useful expression vectors have been developed for this yeast. These vectors are based on an increasing transcription rate in com...

journal_title：BioTechniques

authors： Mimran A,Marbach I,Engelberg D

更新日期：2000-03-01 00:00:00

abstract：:We report a modified method of screening for point mutations using a PCR approach based upon the sensitivity of PCR to the 3' terminus of the primer. This method provides a sensitive screen when using either plasmid DNA or bacterial cell lysates. We have optimized the technique for general use to allow rapid screening...

journal_title：BioTechniques

authors： Major JG Jr

更新日期：1992-01-01 00:00:00

abstract：:Few studies have compared the quantification of mRNA by DNA microarray to the results obtained by reverse transcription PCR (RT-PCR). In this study, mRNA was collected from the healing femoral fracture callus of adult and juvenile rats at various times after fracture. Ten samples were measured by both methods for 26 g...

journal_title：BioTechniques

authors： Etienne W,Meyer MH,Peppers J,Meyer RA Jr

更新日期：2004-04-01 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：: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：:Silencing of mammalian gene expression by RNA interference (RNAi) technology can be achieved using small interfering RNA (siRNA) or short hairpin RNA (shRNA). However, the relative effectiveness of these two approaches is not known. It is also not clear whether gene-specific shRNA transcribed from an RNA polymerase II...

journal_title：BioTechniques

authors： Ling X,Li F

更新日期：2004-03-01 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：:As an improved strategy for producing functional macromolecules by in vitro evolutionary optimization, we propose an automated machine that can process up to 960 samples in parallel. It consists of a 960-well PCR machine with special sealed plastic reaction vessels and appropriate handling devices. We show that the he...

journal_title：BioTechniques

authors： Schober A,Walter NG,Tangen U,Strunk G,Ederhof T,Dapprich J,Eigen M

更新日期：1995-04-01 00:00:00