Fluorescent in situ hybridization employing the conventional NBT/BCIP chromogenic stain.

abstract：:Radiolabeled proteins are used in a variety of laboratory applications as well as in radioimmunotherapy. This review focuses on methods that utilize genetic engineering to introduce exogenous phosphorylation sites into proteins. Protein kinase substrate sites can be introduced into target proteins to serve as tags for...

journal_title：BioTechniques

authors： Clark WA,Izotova L,Philipova D,Wu W,Lin L,Pestka S

更新日期：2002-10-01 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：:Millions of museum specimens are integral to biodiversity studies; however, DNA degradation may limit the ability to obtain DNA sequences. In this study, a degradation analysis model for Lepidoptera specimens was established. Based on this model, we revealed the characteristics of DNA fragment distribution caused by e...

journal_title：BioTechniques

authors： Xu Y,Ren X,Wang H,Wang M,Li G

更新日期：2020-03-01 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 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：:We propose two methods for characterizing the spatio-temporal behavior of cell populations in culture. The first method, image auto-correlation microscopy (IACM), allows us to characterize the variation in the number of objects as a function of time, thus enabling the quantification of the clustering properties of cel...

journal_title：BioTechniques

authors： Bonnet N,Delavoie F,Zahm JM

更新日期：2007-07-01 00:00:00

abstract：:FoLT (formamide low temperature) PCR is a protocol for amplifying DNA directly from whole blood without any preparative steps. Up to 10% (vol/vol) whole blood can be added directly into the tube containing the PCR mixture. There is no need for transfers, centrifugations, pre-boiling or any preparative step. It involve...

journal_title：BioTechniques

authors： Panaccio M,Georgesz M,Lew AM

更新日期：1993-02-01 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：:Plasmid DNA can be purified in 20 min using a three-step CsCl gradient in a vertical tube rotor. Ultracentrifugation with discontinuous gradients yields DNA with purity comparable to that obtained with longer isopycnic separations and with columns. ...

journal_title：BioTechniques

authors： Dorin M,Bornecque CA

更新日期：1995-01-01 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：:We describe a convenient PCR-based protocol for in vitro recombination of homologous genes, thereby minimizing the rate of associated point mutations. High-fidelity recombination conditions were obtained using Vent DNA polymerase, which, in contrast to Taq DNA polymerase, shows significant proofreading activity and ra...

journal_title：BioTechniques

authors： Ninkovic M,Dietrich R,Aral G,Schwienhorst A

更新日期：2001-03-01 00:00:00

abstract：:We have designed and implemented a system to manage whole genome shotgun sequences and whole genome sequence assembly data flow. The Sequence Assembly Manager (SAM) consists primarily of a MySQL relational database and Perl applications designed to easily manipulate and coordinate the analysis of sequence information ...

journal_title：BioTechniques

authors： Warren RL,Butterfield YS,Morin RD,Siddiqui AS,Marra MA,Jones SJ

更新日期：2005-05-01 00:00:00

abstract：:Current gene synthesis methods often incorporate a PCR amplification step in order to yield final material sufficient for resolution from multiple off-products. These amplification steps can cause stochastic sampling effects that propagate errors in gene synthesis or decrease variability when applied to the constructi...

journal_title：BioTechniques

authors： Lee ZB,Firnhaber C,Clarke J,DeDecker BS

更新日期：2015-09-01 00:00:00

abstract：:Human telomerase, a ribonucleoprotein enzyme is known to be associated with immortalized cancer cells but is absent in most normal tissues. Thus, telomerase appears to be an attractive new target for anticancer agents and an important diagnostic marker of human cancers. Here, we describe an improved telomerase assay m...

journal_title：BioTechniques

authors： Sun D,Hurley LH,Von Hoff DD

更新日期：1998-12-01 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：:Piezoelectric dispensing of proteins from borosilicate glass capillaries is a popular method of protein biochip fabrication that offers the advantages of sample recovery and noncontact with the printing substrate. However, little regard has been given to the quantitative aspects of dispensing minute volumes (1 nL or l...

journal_title：BioTechniques

authors： Delehanty JB,Ligler FS

更新日期：2003-02-01 00:00:00

abstract：:Integral membrane G protein-coupled receptors (GPCRs) compose the single most prolific class of drug targets, yet significant functional and structural questions remain unanswered for this superfamily. A primary reason for this gap in understanding arises from the difficulty of forming soluble, monodisperse receptor m...

journal_title：BioTechniques

authors： Leitz AJ,Bayburt TH,Barnakov AN,Springer BA,Sligar SG

更新日期：2006-05-01 00:00:00

abstract：:Genotyping fish larvae is a valuable technique for numerous fields of study. While methods for collecting DNA from early stage larvae have been published, a non-lethal, non-invasive genotyping protocol for hatchlings that is amenable to high-throughput approaches is desirable. Here, we describe a method to individuall...

journal_title：BioTechniques

authors： Espinoza GJ,Poland JM,Bremer JRA

更新日期：2017-10-01 00:00:00

abstract：:A major problem in analyzing forensic casework samples is the presence of genetic material from more than one individual in the material evidence. For instance, in sexual assault cases the evidence (vaginal swabs) usually contains a majority of vaginal epithelial cells and varying amounts of sperm cells from the perpe...

journal_title：BioTechniques

authors： Allen M,Saldeen T,Gyllensten U

更新日期：1995-09-01 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：:Under-agarose chemotaxis has been used previously to assess the ability of neutrophils to respond to gradients of chemoattractant. We have adapted this assay to the chemotactic movement of Dictyostelium amoebae in response to folic acid. Troughs are used instead of wells to increase the area along which the cells can ...

journal_title：BioTechniques

authors： Laevsky G,Knecht DA

更新日期：2001-11-01 00:00:00

abstract：:We describe a new real-time fluorescence video microscope design for capturing intensified images of cells containing dual wavelength "ratio" dyes or multiple dyes. The microscope will perform real-time capture of two separate fluorescence emission images simultaneously, improving the time resolution of spatial distri...

journal_title：BioTechniques

authors： Morris SJ

更新日期：1990-03-01 00:00:00

abstract：:We describe a bead-based, multiplexed, oligonucleotide ligation assay (OLA) performed on the Luminex flow cytometer. Differences between this method and those previously reported include the use of far fewer beads and the use of a universal oligonucleotide for signal detection. These innovations serve to significantly...

journal_title：BioTechniques

authors： Bruse S,Moreau M,Azaro M,Zimmerman R,Brzustowicz L

更新日期：2008-11-01 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：: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：:Oligonucleotide primers used to amplify target DNA regions via PCR should meet certain design criteria to maximize the potential for efficient priming. The Random Oligonucleotide Construction Kit (ROCK), a spreadsheet-based program that runs under Microsoft Excel 97 or later version for Microsoft Windows, was develope...

journal_title：BioTechniques

authors： Strain SR,Chmielewski JG

更新日期：2001-06-01 00:00:00

abstract：:Ditag genome scanning (DGS) uses next-generation DNA sequencing to sequence the ends of ditag fragments produced by restriction enzymes. These sequences are compared to known genome sequences to determine their structure. In order to use DGS for large-scale genome structural studies, we have substantially revised the ...

journal_title：BioTechniques

authors： Jung YC,Xu J,Chen J,Kim Y,Winchester D,Wang SM

更新日期：2009-11-01 00:00:00

abstract：:We describe the application of simple cloning by prolonged overlap extension for multiple site-directed mutagenesis in the same plasmid. We show that it is possible to use this technique with very short PCR templates. The technique is ideally suited for the generation of longer donor DNA sequences for CRISPR/Cas9-medi...

journal_title：BioTechniques

authors： Hejlesen R,Füchtbauer EM

更新日期：2020-06-01 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：:The serum protein transferrin (Tf) is a valuable marker for genetic studies of primates, because it is usually polymorphic, exhibiting as many as 13 allelic forms with high heterozygosity. The standard procedure to detect the different phenotypes requires vertical electrophoresis on polyacrylamide gels for 18 h at 4 d...

journal_title：BioTechniques

authors： Manis GS,Samples NK,Stone WH

更新日期：1993-11-01 00:00:00