Use of signal-distinguishable probes in differential or sequential autoradiography in hybridization analysis.

abstract：:Cytokines are pivotal to a balanced innate or cell-mediated immune response, can be indicative of disease progression and/or resolution, and are being evaluated as therapeutics. There is a need to purify and/or to measure key cytokines rapidly with accuracy, precision, and sensitivity. The current assay technologies, ...

journal_title：BioTechniques

authors： Bai H,Buller RM,Chen N,Boyle MD

更新日期：2002-01-01 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：:Sarah Webb explores how RNA sequencing innovations help scientists tackle the complexity of the transcriptome. ...

journal_title：BioTechniques

authors： Webb S

更新日期：2018-02-01 00:00:00

abstract：:Exposing cells to brief, high-intensity electrical field pulses can lead to the permeabilization of their plasma membranes. This electro-induced permeated state of the cell membrane is reversible and leads to cell fusion; i.e., the electropermeabilized state if fusogenic. The size of cells intended for fusing, however...

journal_title：BioTechniques

authors： Rols MP,Dahhou F,Teissié J

更新日期：1994-10-01 00:00:00

abstract：:S-100B is used in melanoma follow-up. This serum biomarker is also present in adipocytes; therefore, subcutaneous adipocytes trapped in the needle before performing a venipuncture could contaminate the serum. The aim was to study the influence of adipocyte contamination on blood samples used for S-100B analysis, possi...

journal_title：BioTechniques

authors： Damude S,Muller Kobold AC,Bastiaannet E,Kruijff S,Hoekstra HJ,Wevers KP

更新日期：2020-11-01 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：:Scanning conductance microscopy investigations were carried out in air on human chromosomes fixed on pre-fabricated SiO2 surfaces with a backgate. The point of the investigation was to estimate the dielectric constant of fixed human chromosomes in order to use it for microfluidic device optimization. The phase shift c...

journal_title：BioTechniques

authors： Clausen CH,Lange JM,Jensen LB,Shah PJ,Dimaki MI,Svendsen WE

更新日期：2008-02-01 00:00:00

abstract：:We report a straightforward methodology for purification of recombinant proteins by incorporating a short hydrophilic peptide marker segment at their N-termini. A calcium-dependent antibody that reacts primarily with the first three amino acids of this peptide segment was used to affinity purify the fusion proteins in...

journal_title：BioTechniques

authors： Prickett KS,Amberg DC,Hopp TP

更新日期：1989-06-01 00:00:00

abstract：:Sensitive nucleic acid based detection methods such as in situ PCR, in situ RT-PCR and PRINS have great potential in the areas of developmental biology, pathogenesis and diagnostics. However, control of evaporation from in situ reactions is critical to ensure reliable data. Self-Seal Reagent, a component added directl...

journal_title：BioTechniques

authors： Sullivan DE,Bobroski LE,Bagasra O,Finney M

更新日期：1997-08-01 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：:Data are presented illustrating the optimum concentration range of reverse transcription PCR-generated products under 500 bp for accurate base calling with direct automated DNA sequencing. A 357-bp fragment of the human estrogen receptor, which includes the DNA binding domain of the protein, was used as a representati...

journal_title：BioTechniques

authors： Hyder SM,Hu C,Needleman DS,Sonoda Y,Wang XY,Baker VV

更新日期：1994-09-01 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：:The use of affinity-based tools has become invaluable as a platform for basic research and in the development of drugs and diagnostics. Applications include affinity chromatography and affinity tag fusions for efficient purification of proteins as well as methods to probe the protein network interactions on a whole-pr...

journal_title：BioTechniques

authors： Uhlén M

更新日期：2008-04-01 00:00:00

abstract：:Here we describe our progress in the development of the technology of DNA sequencing by primer walking based on the modular primer approach, which eliminates the primer synthesis bottleneck. Modular primers are assembled from 5-mers, 6-mers, or 7-mers selected from a presynthesized library of as few as 1000 oligonucle...

journal_title：BioTechniques

authors： Kotler L,Sobolev I,Ulanovsky L

更新日期：1994-09-01 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：:Several methods and computer programs have been developed for estimating the size of DNA fragments from gel electrophoresis. However, methods are lacking that may facilitate in optimization of gel conditions. In this article, a computer program called DNAOPT is described, which was developed to assist researchers in t...

journal_title：BioTechniques

authors： Raghava GP

更新日期：1995-02-01 00:00:00

abstract：:Studies on the regulation of human milk protein genes and suitable cell culture systems have been limited due to restricted availability of tissue samples from lactating women. Although mammary gland tissue has been available from mammary reduction surgery, studies on tissue-specific expression of milk protein genes r...

journal_title：BioTechniques

authors： Lindquist S,Hansson L,Hernell O,Lönnerdal B,Normark J,Strömquist M,Bergström S

更新日期：1994-10-01 00:00:00

abstract：:We have utilized 5-bromo-2'deoxyuridine (BrdU) substituted DNA as a probe for a number of applications including, principally, for chromosome painting by fluorescence in situ hybridization (FISH) but also for DNA end-labeling to detect apoptotic cell death and for filter hybridization. Br-dUTP was used as a substitute...

journal_title：BioTechniques

authors： Mühlmann-Díaz MC,Dullea RG,Bedford JS

更新日期：1996-07-01 00:00:00

abstract：:The microcomputer-based image analysis system IB-1000 (developed by Indiana Biotech, Highland, IN) for two-dimensional electrophoresis gels has been described previously (9). It allows the user to compare protein spots between two profiles and identify those spots that are commonly shared in both profiles. This report...

journal_title：BioTechniques

authors： Lee C,Sherwood ER,Sensibar JA,Berg LA,Chen YC,Tseng CC

更新日期：1989-04-01 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：:A nonimmune phagemid recombinant antibody fragment (rFab) library was generated with a nominal diversity of 1.16 x 10(7) using the QuikChange Multi Site-Directed Mutagenesis kit. Two degenerate primers spanning the third complementarity-determining region (CDR) loops of the antibody fragment light and heavy chain were...

journal_title：BioTechniques

authors： Kelley LL,Momany C

更新日期：2003-10-01 00:00:00

abstract：:High-throughput approaches for gene cloning and expression require the development of new nonstandard tools for molecular biologists and biochemists. We introduce a Web-based tool to design primers specifically for the generation of expression clones for both laboratory-scale and high-throughput projects. The applicat...

journal_title：BioTechniques

authors： Yoon JR,Laible PD,Gu M,Scott HN,Collart FR

更新日期：2002-12-01 00:00:00

abstract：:We describe a method using an inexpensive craft glue to routinely isolate specific areas of tissue as small as 1 mm2 from paraffin sections. The tissue may be digested to release nucleic acid suitable for PCR or reverse transcription PCR. The use of this procedure obviates the requirement for manual microdissection or...

journal_title：BioTechniques

authors： Turbett GR,Barnett TC,Dillon EK,Sellner LN

更新日期：1996-05-01 00:00:00

abstract：:Here we describe an amplification method for global transcript analysis. The strategy relies on amplification of cDNA tags (signature tags) achieved by random fragmentation of the cDNAs to short tags of similar length, isolation of the 3' ends and then PCR amplification of the 3'-end signature tag population. This met...

journal_title：BioTechniques

authors： Sievertzon M,Agaton C,Nilsson P,Lundeberg J

更新日期：2004-02-01 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：:[Formula: see text] Known to be a sturdy weapon in a scientist's arsenal, how has the gene editing tool CRISPR been applied in the fight against COVID-19? ...

journal_title：BioTechniques

authors： Straiton J

更新日期：2020-11-01 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：:As a strategy to purify recombinant murine Interleukin (IL)-18, we cloned the mature coding region of this protein into the pFLAG-1 expression system. The intent was to use the FLAG peptide "tag" as an amino terminal addition to IL-18 so that purification of this fusion protein (FLAG-IL-18) on anti-FLAG antibody affin...

journal_title：BioTechniques

authors： Elhofy A,Bost KL

更新日期：1998-09-01 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：: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