A method for visualizing fluorescence of flavonoid therapeutics in vivo in the model eukaryote Dictyostelium discoideum.

abstract：:The use of reverse transcription (RT) PCR for relative quantitation of gene transcripts relies on the reproducibility of the individual RT, PCR and product measurement steps. Semi-competitive RT-PCR (RT-cPCR) uses an internal competitor template in the PCR step to improve quantitation. We have surveyed the reproducibi...

journal_title：BioTechniques

authors： Hall LL,Bicknell GR,Primrose L,Pringle JH,Shaw JA,Furness PN

更新日期：1998-04-01 00:00:00

abstract：:We have developed a rapid, sensitive, and quantitative 96-well microplate-based nonradioactive immunoprecipitation/kinase assay to evaluate mitogen-activated protein kinase (MAPK) activity. Three quantitative nonradioactive imunoprecipitation/kinase assays of MAPK were demonstrated on a 96-well microplate coated with ...

journal_title：BioTechniques

authors： Zhang L,Uder S,Juehne T,Brizzard B,Song K

更新日期：2002-02-01 00:00:00

abstract：:The colony formation assay (CFA) is the gold standard for measuring the effects of cytotoxic agents on cancer cells in vitro; however, in its traditional 6-well format, it is a time-consuming assay, particularly when evaluating combination therapies. In the interest of increased efficiency, the 6-well CFA was converte...

journal_title：BioTechniques

authors： Katz D,Ito E,Lau KS,Mocanu JD,Bastianutto C,Schimmer AD,Liu FF

更新日期：2008-02-01 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：: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：:We describe here a simple and efficient transfection method for transient expression of cloned genes in cell lines and primary cultured cells. The method involves the use of DEAE-dextran to target DNA to the cellular endocytotic pathway and the use of a human adenovirus to ensure efficient lysis of endosomal vesicles....

journal_title：BioTechniques

authors： Forsayeth JR,Garcia PD

更新日期：1994-08-01 00:00:00

abstract：:Many DNA binding proteins are known to regulate gene expression. When that binding is altered, a disease state can result. A common method for measuring DNA binding, namely electrophoretic mobility shift assay (EMSA) is often used but it is not amenable to rapid screening of many samples. As an alternative method, we ...

journal_title：BioTechniques

authors： Gal S,Cook JR,Howells L

更新日期：2006-09-01 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：:Previous work from this laboratory has shown that immunoporation has the potential for the selective transfection of a range of different animal cells based on their immunological identity. The unique ability of immunoporation to target cells for transfection combined with the high efficiency of transfection and the h...

journal_title：BioTechniques

authors： Tzavelas C,Bildirici L,Rickwood D

更新日期：2004-08-01 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：:The well-characterized cell line Chinese hamster ovary (CHO) has been used to produce numerous biopharmaceuticals and is an important tool for basic research. However, introducing foreign DNA into specially modified CHO cells such as DG44 and Lec 3.2.8.1 can sometimes be an arduous process. Here we show that the Flp-i...

journal_title：BioTechniques

authors： Soler DC,Young AE,Vahedi-Faridi A,McCormick TS

更新日期：2018-07-01 00:00:00

abstract：:The use of quantitative PCR is recommended to monitor the level of residual hematological malignancies. The proposed multiplex IgH/ras PCR uses a co-amplification of the clonal CDR3 rearrangement of the immunoglobulin heavy chain gene (IgH) as a disease marker and a segment of the Hras 1 gene containing codon 61 (ras)...

journal_title：BioTechniques

authors： Slavícková A,Ullmannová V,Klener P

更新日期：2000-04-01 00:00:00

abstract：:The nematode Caenorhabditis elegans is an important model animal for biological research. Currently, transgenic C. elegans strains are mainly generated by injecting DNA encoding a gene of interest, in combination with a reporter gene, into the gonad. With this approach, the interpretation of negative results, such as ...

journal_title：BioTechniques

authors： Li D,Wang M

更新日期：2012-03-01 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：:We previously reported that cervicovaginal lavages (CVL) contain a factor that enhances the replication of human immunodeficiency virus (HIV) by increasing virus transcription in T cells and monocytoid cells. This factor was named the HIV-inducing factor (HIF). To determine the molecular mass of HIF, we adapted a blot...

journal_title：BioTechniques

authors： Hashemi FB,Spear GT,Madsen L,Mollenhauer J

更新日期：2000-03-01 00:00:00

abstract：:A nascent protein remains in a complex with its ribosome and mRNA if the stop codon is deleted from the mRNA. In the same manner, mRNA forms a stable complex with DNA if the transcription termination is blocked. In principle, if both mRNA translation and DNA transcription termination are prevented, the protein should ...

journal_title：BioTechniques

authors： Liu S,Shivashankar GV,Sano T,Libchaber A

更新日期：2000-10-01 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：:To increase the reproducibility and to reduce the false positives in the initial mRNA differential display, modified long composite primers were developed based on both mRNA differential display and RNA arbitrarily primed PCR fingerprinting methods. Ten-base nucleotides were added at the 5' ends of the primers used in...

journal_title：BioTechniques

authors： Zhao S,Ooi SL,Pardee AB

更新日期：1995-05-01 00:00:00

abstract：:The increase of information in biology makes it difficult for researchers in any field to keep current with the literature. The MEDLINE database of scientific abstracts can be quickly scanned using electronic mechanisms. Potentially interesting abstracts can be selected by matching words joined by Boolean operators. H...

journal_title：BioTechniques

authors： Perez-Iratxeta C,Keer HS,Bork P,Andrade MA

更新日期：2002-06-01 00:00:00

abstract：:In this report we present a rapid and inexpensive PCR-based method to screen recombinant DNA libraries. The efficiency of this method was demonstrated by the isolation of clones of interest from three different libraries using different vector systems. This method is nonradioactive and makes it easier to handle a larg...

journal_title：BioTechniques

authors： Amaravadi L,King MW

更新日期：1994-01-01 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：: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：:A fully automated nucleic acid analysis system is described, which offers positive sample identification, improved sensitivity and reduced user interaction compared to conventional techniques. The system relies on the sequence-specific capture of DNA onto solid-phase particles, confirming product identity without the ...

journal_title：BioTechniques

authors： Brown A,Akinsanya AA,Barker SJ,Brophy M,Dobb AK,Doyle SM,Hudson IR,Minter SJ,Wraith MJ,Oultram JD

更新日期：1999-07-01 00:00:00

abstract：:Transfection of suspension cells has proven to be very difficult using conventional methods. Here, we present a simple and time-saving new transfection protocol wherein cell culture plates coated with chicken egg white are seeded with suspension cells prior to transfection. Our results demonstrate that coupling egg wh...

journal_title：BioTechniques

authors： Basiouni S,Fuhrmann H,Schumann J

更新日期：2012-08-01 00:00:00

abstract：:We have demonstrated that efficient polymerase chain reaction amplifications from chromosomal DNA can be carried out using whole bacterial cells as the starting material. Cells from the liquid or solid cultures can be used directly, without any pre-treatment, thus eliminating the need for DNA isolation. ...

journal_title：BioTechniques

authors： Joshi AK,Baichwal V,Ames GF

更新日期：1991-01-01 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：: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：:A method for cloning single-stranded oligonucleotides in a plasmid vector has been developed. The method relies on ligation of the oligonucleotide into suitable restriction enzyme sites of the cloning vector such that the site at the 5' end has a 5' overhang [for example, a Bgl II site (A decreases GATCT)], and the si...

journal_title：BioTechniques

authors： Mounts P,Wu TC,Peden K

更新日期：1989-04-01 00:00:00

abstract：:This report describes a new method for labeling PCR-generated short length (60-120 bp) double-stranded DNA fragments for use as hybridization probes. The method utilizes gene-specific primers identical to those for PCR generation of non-radioactive DNA fragments. Radioactive probes are synthesized by Taq DNA polymeras...

journal_title：BioTechniques

authors： Mizobuchi M,Frohman LA

更新日期：1992-03-01 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