SOP3: a web-based tool for selection of oligonucleotide primers for single nucleotide polymorphism analysis by Pyrosequencing.

abstract：:Chromosome walking in mammalian DNA by vectorette PCR is not always very specific, and the walks have been limited to distances <1 kb. To improve the method, we have designed new vectorettes, which unlike the currently used ones have very little repetitive sequences or homology with known DNA sequences of various orig...

journal_title：BioTechniques

authors： Laitinen J,Räkköläinen T,Hölttä E

更新日期：2004-10-01 00:00:00

abstract：:3D printed biomaterials are increasingly used in cell cultures and drug screens. Given the ease of creating artificial tissues, will this technique revolutionize biomedicine and organ implants in the future? ...

journal_title：BioTechniques

authors： Prabhune M

更新日期：2018-03-01 00:00:00

abstract：:Substantial technological advances in proteomics and related computational science have been made in the past few years. These advances overcome in part the complexity and heterogeneity of the human proteome, permitting quantitative analysis and identification of protein changes associated with tumor development. Here...

journal_title：BioTechniques

authors： Faca V,Krasnoselsky A,Hanash S

更新日期：2007-09-01 00:00:00

abstract：:A variety of technical errors have arisen in data analysis when using cDNA or oligonucleotide microarrays. One of the most insidious problems is the saturation of the hybridization signal of high-abundant transcripts. This problem arises from the truncation of the laser fluorescence signal. When the hybridization sign...

journal_title：BioTechniques

authors： Hsiao LL,Jensen RV,Yoshida T,Clark KE,Blumenstock JE,Gullans SR

更新日期：2002-02-01 00:00:00

abstract：:Ribosome profiling is a powerful tool for characterizing in vivo protein translation at the genome scale, with multiple applications ranging from detailed molecular mechanisms to systems-level predictive modeling. Though highly effective, this intricate technique has yet to become widely used in the microbial research...

journal_title：BioTechniques

authors： Latif H,Szubin R,Tan J,Brunk E,Lechner A,Zengler K,Palsson BO

更新日期：2015-06-01 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：:We report here an improved method for analyzing protein surface expression utilizing a cold-adapted trypsin. Preservation of activity of the enzyme at 0-4°C permits modification of the protease method of surface analysis to temperatures at which trafficking of mammalian plasmalemmal proteins is blocked. This is an imp...

journal_title：BioTechniques

authors： Ahmad F,Coleman SK,Kaila K,Blaesse P

更新日期：2011-04-01 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 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：: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：:Screening methods for chemotherapeutic agents usually rely on the cytotoxic properties of the drugs. However, agents that inhibit invasion may have more efficacy and cause fewer side effects. Various cellular invasion assays have been used to evaluate these types of compounds, including the modified Boyden chamber, mo...

journal_title：BioTechniques

authors： Sasaki CY,Passaniti A

更新日期：1998-06-01 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：: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：: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：:Cell-imaging approaches using new laser-based technologies have a wide applicability to thefields of pathology and cell biology. Here, we present the application of several of these techniques, including confocal scanning laser microscopy (CSLM), laser scanning cytometry (LSC), and laser capture microdissection (LCM),...

journal_title：BioTechniques

authors： Taatjes DJ,Palmer CJ,Pantano C,Hoffmann SB,Cummins A,Mossman BT

更新日期：2001-10-01 00:00:00

abstract：:A method for the isolation of total cytoplasmic RNA and high molecular weight DNA from the same cells is described. Cells are gently lysed with NP40 in the presence of vanadyl ribonucleoside complex and the nuclei pelleted by centrifugation. RNA is purified by phenol/CHCl3 extraction of the lysate supernatant followed...

journal_title：BioTechniques

authors： Nicolaides NC,Stoeckert CJ Jr

更新日期：1990-02-01 00:00:00

abstract：:Cysts of free-living protozoa have an impact on the ecology and epidemiology of bacteria because they may act as a transmission vector or shelter the bacteria against hash environmental conditions. Detection and localization of intracystic bacteria and examination of the en- and excystment dynamics is a major challeng...

journal_title：BioTechniques

authors： Lambrecht E,Baré J,Claeys M,Chavatte N,Bert W,Sabbe K,Houf K

更新日期：2015-04-01 00:00:00

abstract：:Few studies have focused on the significance of ras protein levels in human malignancy, in part because of the inherent difficulty in quantitation of the ras gene product. We have developed a method for the enzymatic determination of the ras gene product and have used this method for the quantitation of ras gene produ...

journal_title：BioTechniques

authors： Gutheil JC,Mane SM,Bass KA,Lee EJ,Needleman SW

更新日期：1990-08-01 00:00:00

abstract：:A method is described for preparing mutants with multiple, site-directed mutations by ordered coupling of PCR-generated fragments catalyzed by a thermostable DNA ligase. Annealing of the sense strands of the fragments to a single-stranded (antisense) template created a full-length sense strand leaving only nicks that ...

journal_title：BioTechniques

authors： Rouwendal GJ,Wolbert EJ,Zwiers LH,Springer J

更新日期：1993-07-01 00:00:00

abstract：:Genomic analysis of circulating, cell-free DNA (cfDNA) is being used extensively for molecular diagnostics. Many approaches rely on the construction of cfDNA genomic libraries, targeted retrieval of specific genomic regions and analysis by next-generation DNA sequencing. Several steps during sample preparation require...

journal_title：BioTechniques

authors： Raymond CK

更新日期：2019-10-01 00:00:00

abstract：:Identification of antimitotic molecules that affect tubulin dynamics is a multistep procedure. It includes in vitro tubulin polymerization assay, studies of a cell cycle effect, and general cytotoxicity assessment. To simplify this lengthy screening protocol, we have introduced and validated an assay system based on t...

journal_title：BioTechniques

authors： Semenova MN,Kiselyov A,Semenov VV

更新日期：2006-06-01 00:00:00

abstract：:Primary human fibroblasts and a series of cell lines (A549, BNL CL.2, H225, NIH 3T3 and Rat-1) are efficiently transfected by using positively charged complexes of plasmid DNA and transferrin-polylysine or polylysine in the presence of glycerol (1 molar to 1.8 molar, depending on the cell type). An increase in gene ex...

journal_title：BioTechniques

authors： Zauner W,Kichler A,Schmidt W,Sinski A,Wagner E

更新日期：1996-05-01 00:00:00

abstract：:The yeast two-hybrid system has been used to characterize many protein-protein interactions. A two-hybrid system for E. coli was constructed in which one hybrid protein bound to a specific DNA site recruits another to an adjacent DNA binding site. The first hybrid comprises a test protein, the bait, fused to a chimeri...

journal_title：BioTechniques

authors： Hays LB,Chen YS,Hu JC

更新日期：2000-08-01 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：:A modification of the TRI Reagent procedure has been elaborated for isolation of RNA from polysaccharide- and proteoglycan-rich material. In the modified procedure, RNA is precipitated from the aqueous phase by the combined action of isopropanol and a high-salt concentration. Under these conditions, RNA is effectively...

journal_title：BioTechniques

authors： Chomczynski P,Mackey K

更新日期：1995-12-01 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：: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：:A magnetic bead-based system for DNA isolation utilizing monodisperse beads was tested with the aim of producing a general approach for PCR-ready DNA. This commercially available system was originally designed for isolating PCR-ready DNA from human whole blood. We tested diverse organisms belonging to the major groups...

journal_title：BioTechniques

authors： Rudi K,Kroken M,Dahlberg OJ,Deggerdal A,Jakobsen KS,Larsen F

更新日期：1997-03-01 00:00:00

abstract：:Large genes present particular cloning difficulties, especially when expressed at relatively low levels. We describe a novel method, termed 3' rapid amplification of cDNA ends (RACE) walking, for the rapid determination of unknown 3' flanking sequence of a large cDNA. The technique is a derivative of the anchored PCR ...

journal_title：BioTechniques

authors： Park DJ,Pask AJ,Renfree MB,Graves JA

更新日期：2003-04-01 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