Monitoring helicase activity with molecular beacons.

abstract：:A method allowing the evaluation of the structure and the calculation of the volume of a biofilm, using an optical microscope, is proposed based on the linear relation between the intensity of a pixel in biofilm images grabbed on the x-y plane and the corresponding number of cells in the z direction, which allows the ...

journal_title：BioTechniques

authors： de Carvalho CC,da Fonseca MM

更新日期：2007-05-01 00:00:00

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：:A new procedure is described for the generation of high-titer, helper-free retrovirus vectors employing receptor-mediated, adenovirus-augmented transfection into a standard packaging cell line. Viral titers are increased 30-fold to 100-fold in transiently (> 10(5) infectious units per mL) and stable (> 10(7) infectiou...

journal_title：BioTechniques

authors： von Rüden T,Stingl L,Cotten M,Wagner E,Zatloukal K

更新日期：1995-03-01 00:00:00

abstract：:We describe a rapid, simple and inexpensive method for the isolation of DNA from blood clots suited for use in PCR. Our method is based on the lysing and nuclease-inactivating properties of guanidine thiocyanate together with the nucleic acid-binding properties of silica particles. Isolated DNA can be used for in vitr...

journal_title：BioTechniques

authors： Zeillinger R,Schneeberger C,Speiser P,Kury F

更新日期：1993-02-01 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：:An improved Huffman coding method for information storage in DNA is described. The method entails the utilization of modified unambiguous base assignment that enables efficient coding of characters. A plasmid-based library with efficient and reliable information retrieval and assembly with uniquely designed primers is...

journal_title：BioTechniques

authors： Ailenberg M,Rotstein O

更新日期：2009-09-01 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：:Loop-mediated isothermal amplification (LAMP), a novel gene amplification method, enables the synthesis of larger amounts of both DNA and a visible byproduct--namely, magnesium pyrophosphate--without thermal cycling. A positive reaction is indicated by the turbidity of the reaction solution or the color change after a...

journal_title：BioTechniques

authors： Goto M,Honda E,Ogura A,Nomoto A,Hanaki K

更新日期：2009-03-01 00:00:00

abstract：:Polyethyleneimine (PEI) is a flocculent that is widely used in the downstream purification of monoclonal antibodies. It is an in-process residual that is carried through the drug purification process and strongly inhibits residual DNA quantitation by real-time quantitative PCR assay. Very high sample dilutions (e.g., ...

journal_title：BioTechniques

authors： Zhang SM,Roberts M,Jones M,Zeitler J,Kilby G,Liu A,White JR

更新日期：2020-06-01 00:00:00

abstract：:Bionanotechnology is an emerging field in nanotechnology. In general, it uses concepts from chemistry, biochemistry, and molecular biology to identify components and processes for the construction of self-assembling materials and devices. Distant goals of the science of bionanotechnology range from developing programm...

journal_title：BioTechniques

authors： Clark J,Singer EM,Korns DR,Smith SS

更新日期：2004-06-01 00:00:00

abstract：:Immunofluorescence quantification of γH2AX foci is a powerful approach to quantify DNA double-strand breaks induced by cancer therapy or accidental exposure to ionizing radiation. Here we report a modification to the γH2AX immunofluorescence labeling method, whereby cells are stained in-solution before being spotted a...

journal_title：BioTechniques

authors： Johansson P,Muslimovic A,Hultborn R,Fernström E,Hammarsten O

更新日期：2011-09-01 00:00:00

abstract：:MicroRNAs (miRNAs) are ~22-nucleotide-long small non-coding RNAs that regulate the expression of protein-coding genes by base pairing to partially complementary target sites, preferentially located in the 3´ untranslated region (UTR) of target mRNAs. The expression and function of miRNAs have been extensively studied ...

journal_title：BioTechniques

authors： Persson H,Søkilde R,Pirona AC,Rovira C

更新日期：2017-08-01 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：:The Sequence Analysis Workshop (SAW) is an interactive program for sequence analysis of immunoglobulin variable domains. Sequences for SAW can be obtained from GenBank or from a standard text file. SAW can compare a variable domain to as many as 100 different sequences, calculate the extent of homology, sort the seque...

journal_title：BioTechniques

authors： Elgavish RA,Schroeder HW Jr

更新日期：1993-12-01 00:00:00

abstract：:A technique, Replacement PCR Mutagenesis, was developed to replace one immunoglobulin variable region (V) in a M13 phage cassette with a different, homologous V. This allows the use of the same mutagenesis and subsequent expression vectors for many V regions or V segments. The method combines PCR of V fragments and in...

journal_title：BioTechniques

authors： Near RI

更新日期：1992-01-01 00:00:00

abstract：:We have developed a novel vector pTCS, as a tool for efficient selection of open reading frame (ORF)-containing inserts. In pTCS clones containing an insert with an ORF a downstream marker gene (immE3, conferring resistance to colicin) is activated via translational coupling with the insert, and transformed cells can ...

journal_title：BioTechniques

authors： Ohashi-Kunihiro S,Yohda M,Masaki H,Machida M

更新日期：2007-12-01 00:00:00

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：: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：:Human multiple tissue Western (MTW) blots are premade immunoblots prepared using proteins isolated from adult human tissue. The proteins are isolated from whole tissue homogenates under conditions designed to minimize proteolysis and to ensure maximal representation of tissue-specific proteins. Sodium dodecyl sulfate ...

journal_title：BioTechniques

authors： Kain SR,Mai K,Sinai P

更新日期：1994-11-01 00:00:00

abstract：:A procedure for amplification by PCR of reproducible allele markers for amplified fragment length polymorphism (Amp-FLP) analysis is presented. We have prepared markers for the allelic products of the VNTR loci D1S80 (MCT118) and D17S30 (YNZ22) and for the hypervariable VNTR locus close to the 3' end of the apolipopro...

journal_title：BioTechniques

authors： Sajantila A,Puomilahti S,Johnsson V,Ehnholm C

更新日期：1992-01-01 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：:Cycling probe technology (CPT) is a simple signal amplification method for the detection of specific target DNA sequences. CPT uses a chimeric DNA-RNA-DNA probe that is cut by RNase H when bound to its complementary target sequence. In this study, a hybridization assay was developed to detect biotinylated CPT products...

journal_title：BioTechniques

authors： Warnon S,Zammatteo N,Alexandre I,Hans C,Remacle J

更新日期：2000-06-01 00:00:00

abstract：:We report a robust method for studying en masse changes in translation using cDNA arrays. The relative distribution of messenger RNAs (mRNAs) along polysome gradients was monitored by performing cDNA array analysis of each gradient fraction and quantifying the mRNA translational status by regression analysis. Using th...

journal_title：BioTechniques

authors： Mazan-Mamczarz K,Kawai T,Martindale JL,Gorospe M

更新日期：2005-07-01 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：:Changes in cardiac structure that depart from normal have generally been termed "remodeling". Assessment of ventricular remodeling at the cellular level should include measurement of myocyte dimensions. A well-established and reliable method to assess myocyte remodeling uses isolated cells and the Coulter Counter/Chan...

journal_title：BioTechniques

authors： Said S,Tamura T,Gerdes AM

更新日期：1998-09-01 00:00:00

abstract：:Antibodies specific to avian myeloblastosis virus envelope glycoprotein gp80 were raised. Immunoliposomes were prepared using anti-avian myeloblastosis virus envelope glycoprotein gp80 antibody. The antibody was palmitoylated to facilitate its incorporation into lipid bilayers of liposomes. The fluorescence emission s...

journal_title：BioTechniques

authors： Kalvakolanu DV,Abraham A

更新日期：1991-08-01 00:00:00

abstract：:We have conducted a systematic evaluation of the calculated molecular properties of compounds in clinical development and have found that the development process selects for compounds that have certain computed physical properties. In particular, as the stage of development progresses, compounds that are advanced have...

journal_title：BioTechniques

authors： Blake JF

更新日期：2003-06-01 00:00:00

abstract：:Manipulating gene expression in primary neurons has been a goal for many scientists for over 20 years. Vertebrate central nervous system neurons are classically difficult to transfect. Most lipid reagents are inefficient and toxic to the cells, and time-consuming methods such as viral infections are often required to ...

journal_title：BioTechniques

authors： Buchser WJ,Pardinas JR,Shi Y,Bixby JL,Lemmon VP

更新日期：2006-11-01 00:00:00

abstract：:Specific amplification of nucleic acid sequences by PCR has been extensively used for the detection of gene rearrangements and gene expression. Although successful amplification of DNA sequences has been carried out with DNA prepared from formalin-fixed, paraffin-embedded (FFPE) tissues, there are only a few reports r...

journal_title：BioTechniques

authors： Finke J,Fritzen R,Ternes P,Lange W,Dölken G

更新日期：1993-03-01 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