Application of 5-bromo-2'deoxyuridine as a label for in situ hybridization in chromosome microdissection and painting, and 3' OH DNA end labeling for apoptosis.

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 for biotin or digoxigenin-dUTP in probe labeling techniques, such as random priming, nick translation, end-labeling or PCR. An especially useful application is that it may be incorporated into probe DNA while cells or plasmids in bacteria are growing in the presence of BrdU. This can be particularly advantageous when large quantities of probe are needed, since the cost per mole of digoxigenin-dUTP or biotin-dUTP is nearly 1000 times that of Br-dUTP. Also, if probe is prepared by growth in BrdU, the difference in cost to prepare equal quantities of labeled DNA is more than 10,000 times greater for biotin-dUTP.

journal_name

Biotechniques

journal_title

BioTechniques

authors

Mühlmann-Díaz MC,Dullea RG,Bedford JS

subject

Has Abstract

pub_date

1996-07-01 00:00:00

pages

82-6

issue

1

eissn

0736-6205

issn

1940-9818

journal_volume

21

pub_type

  • The use of synthetic peptide combinatorial libraries for the identification of bioactive peptides.

    abstract::The systematic preparation of synthetic peptide combinatorial libraries (SPCLs), each composed of tens of millions of peptides that can be screened in existing diagnostically or pharmacologically relevant in vitro assay systems, is reviewed. The identification of optimal peptide sequences has been achieved through the...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Houghten RA,Appel JR,Blondelle SE,Cuervo JH,Dooley CT,Pinilla C

    更新日期:1992-09-01 00:00:00

  • Nonradioactive assay for sequence-specific DNA binding proteins.

    abstract::A nonradioactive functional assay was developed to quantitate DNA binding proteins. The assay was designed to allow the use of 96-well microplates for high sample throughput. We show that the assay can measure sequence-specific DNA binding of purified proteins as well as DNA binding activity present in whole cell extr...

    journal_title:BioTechniques

    pub_type:

    doi:

    authors: Gubler ML,Abarzúa P

    更新日期:1995-06-01 00:00:00

  • Impact of RNA extraction from limited samples on microarray results.

    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

    pub_type:

    doi:10.2144/03355st04

    authors: Ojaniemi H,Evengard B,Lee DR,Unger ER,Vernon SD

    更新日期:2003-11-01 00:00:00

  • Evaluation of DNA degradation and establishment of a degradation analysis model for Lepidoptera specimens.

    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

    pub_type: 杂志文章

    doi:10.2144/btn-2019-0166

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

    更新日期:2020-03-01 00:00:00

  • Staining and embedding of human chromosomes for 3-d serial block-face scanning electron microscopy.

    abstract::The high-order structure of human chromosomes is an important biological question that is still under investigation. Studies have been done on imaging human mitotic chromosomes using mostly 2-D microscopy methods. To image micron-sized human chromosomes in 3-D, we developed a procedure for preparing samples for serial...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/000114236

    authors: Yusuf M,Chen B,Hashimoto T,Estandarte AK,Thompson G,Robinson I

    更新日期:2014-12-01 00:00:00

  • A simple microfluidic device for live cell imaging of Arabidopsis cotyledons, leaves, and seedlings.

    abstract::One of the challenges of performing live-cell imaging in plants is establishing a system for securing the sample during imaging that allows for the rapid addition of treatments. Here we report how a commercially available device called a HybriWell™ can be repurposed to create an imaging chamber suitable for Arabidopsi...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/btn-2018-0044

    authors: Vang S,Seitz K,Krysan PJ

    更新日期:2018-06-01 00:00:00

  • Rapid purification of synthetic oligonucleotides: a convenient alternative to high-performance liquid chromatography and polyacrylamide gel electrophoresis.

    abstract::A new method has been developed to purify and detritylate milligram amounts of synthetic oligonucleotides. Dimethoxytrityl oligonucleotides from 15 to 100 nucleotides in length are applied in triethylammonium acetate or concentrated ammonium hydroxide to a disposable chromatographic cartridge, the NENSORB PREP Nucleic...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Johnson BA,McClain SG,Doran ER,Tice G,Kirsch MA

    更新日期:1990-04-01 00:00:00

  • Surrogate marker for predicting the virus binding of urogenital cancer cells during adenovirus-based gene therapy.

    abstract::Virus uptake is the first rate-limiting step for the successful gene delivery of any virus-based gene therapy. For adenovirus-based gene therapy, the expression levels of the adenovirus receptor--coxsackievirus and adenovirus receptor (CAR)--play an important role in dictating gene delivery. We have observed a wide sp...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/03351dd02

    authors: Lai YJ,Pong RC,McConnell JD,Hsieh JT

    更新日期:2003-07-01 00:00:00

  • An effective, low-cost method for achieving and maintaining hypoxia during cell culture studies.

    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

    pub_type: 杂志文章

    doi:10.2144/000114341

    authors: Bakmiwewa SM,Heng B,Guillemin GJ,Ball HJ,Hunt NH

    更新日期:2015-10-01 00:00:00

  • A rapid and simple bead-bashing-based method for genomic DNA extraction from mammalian tissue.

    abstract::Conventional genomic DNA (gDNA) extraction methods can take hours to complete, may require fume hoods and represent the most time-consuming step in many gDNA-based molecular assays. We systematically optimized a bead bashing-based (BBB) approach for rapid gDNA extraction without the need for a fume hood. Human tissue ...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/btn-2019-0172

    authors: Wei S,Levy B,Hoffman N,Cujar C,Rodney-Sandy R,Wapner R,D'Alton M,Williams Z

    更新日期:2020-05-01 00:00:00

  • The era of 'omics unlimited.

    abstract::Determining the primary sequences of informational macromolecules is no longer a limiting factor for our ability to completely understand the biological functioning of cells and organisms. Similarly, our understanding of transcriptional regulation (transcriptomics) has been greatly enhanced by the availability of micr...

    journal_title:BioTechniques

    pub_type: 杂志文章,评审

    doi:10.2144/000113137

    authors: Kandpal R,Saviola B,Felton J

    更新日期:2009-04-01 00:00:00

  • DIRECTING DRUG EVOLUTION.

    abstract::Directed evolution is poised to change small molecule discovery and provide greater access to "drug space." Sarah Webb looks into the evolving drug discovery landscape. ...

    journal_title:BioTechniques

    pub_type: 新闻

    doi:10.2144/000114616

    authors: Webb S Ph D

    更新日期:2017-12-01 00:00:00

  • PBXL-1: a new fluorochrome applied to detection of proteins on membranes.

    abstract::An easy, sensitive and direct fluorescent immunodetection method for proteins is described using the new fluorochrome PBXL-1 imaged with the FMBIO II Laser Scanning Imaging System. PBXL-1 is derived from a protein supra-molecular complex that contains a large number of chromophores. This complex, the phycobilisome, is...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/99263pf02

    authors: Morseman JP,Moss MW,Zoha SJ,Allnutt FC

    更新日期:1999-03-01 00:00:00

  • Preparation of amino acid mixtures for cell-free expression systems.

    abstract::Here we present a procedure for preparing amino acid mixtures--having both the desired composition and a physiological pH--at high concentrations for cell-free expression systems. Up to 2.1 mg/mL of active protein was synthesized in batch mode reactions with an all Escherichia coli cell-free expression system. Our met...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/000114249

    authors: Caschera F,Noireaux V

    更新日期:2015-01-01 00:00:00

  • Direct RT-PCR amplification of mRNA supported on membranes.

    abstract::We describe a simple and efficient technique that facilitates the amplification of specific mRNA for cloning and sequencing purposes. An mRNA bound to a small piece of membrane filter is used as a template to synthesize complementary DNA. The product of this reaction is then transferred to a new tube and amplified usi...

    journal_title:BioTechniques

    pub_type:

    doi:

    authors: Ruiz A,Bok D

    更新日期:1993-11-01 00:00:00

  • Detection of bacterial mRNA using polymerase chain reaction.

    abstract::A method was developed for the detection of bacterial mRNAs using reverse transcriptase followed by the polymerase chain reaction (PCR) and Southern blot analysis. The method involves brief inhibition of protein synthesis with chloramphenicol, followed by reverse transcription, PCR amplification of cDNA and Southern b...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Mahbubani MH,Bej AK,Miller RD,Atlas RM,DiCesare JL,Haff LA

    更新日期:1991-01-01 00:00:00

  • SAW: a graphical user interface for the analysis of immunoglobulin variable domain sequences.

    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

    pub_type:

    doi:

    authors: Elgavish RA,Schroeder HW Jr

    更新日期:1993-12-01 00:00:00

  • Pyrophosphorolysis-activated polymerization (PAP): application to allele-specific amplification.

    abstract::To measure mutation load or to detect minimal residual disease, a robust method for identifying one mutant allele in the range of 10(6)-10(9) wild-type alleles would be advantageous. Herein, we present evidence that pyrophosphorolysis-activated polymerization (PAP) has the potential to provide a highly specific and ro...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/00295rr03

    authors: Liu Q,Sommer SS

    更新日期:2000-11-01 00:00:00

  • Genotyping live fish larvae: Non-lethal and noninvasive DNA isolation from 3-5 day old hatchlings.

    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

    pub_type: 杂志文章

    doi:10.2144/000114598

    authors: Espinoza GJ,Poland JM,Bremer JRA

    更新日期:2017-10-01 00:00:00

  • DNA sequencing: modular primers for automated walking.

    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

    pub_type: 杂志文章

    doi:

    authors: Kotler L,Sobolev I,Ulanovsky L

    更新日期:1994-09-01 00:00:00

  • A DNA cassette containing a trimerized SV40 polyadenylation signal which efficiently blocks spurious plasmid-initiated transcription.

    abstract::A head-to-tail trimer of the SV40 Bcl I-Bam H1 DNA fragment, specifying polyadenylation of RNA transcripts, was cloned as a cassette flanked by multiple restriction sites. Insertion of the trimer into several expression vectors efficiently prevented spurious expression of reporter genes resulting from transcriptional ...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Maxwell IH,Harrison GS,Wood WM,Maxwell F

    更新日期:1989-03-01 00:00:00

  • Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material.

    abstract::Procedures utilizing Chelex 100 chelating resin have been developed for extracting DNA from forensic-type samples for use with the PCR. The procedures are simple, rapid, involve no organic solvents and do not require multiple tube transfers for most types of samples. The extraction of DNA from semen and very small blo...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Walsh PS,Metzger DA,Higuchi R

    更新日期:1991-04-01 00:00:00

  • Continuous fluorescence monitoring of rapid cycle DNA amplification.

    abstract::Rapid cycle DNA amplification was continuously monitored by three different fluorescence techniques. Fluorescence was monitored by (i) the double-strand-specific dye SYBR Green I, (ii) a decrease in fluorescein quenching by rhodamine after exonuclease cleavage of a dual-labeled hydrolysis probe and (iii) resonance ene...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/97221bi01

    authors: Wittwer CT,Herrmann MG,Moss AA,Rasmussen RP

    更新日期:1997-01-01 00:00:00

  • Quantitation of PCR products with chemiluminescence.

    abstract::Quantitative PCR and reverse transcription PCR (RT-PCR) are widely used in biomedical, industrial and other research applications to determine the number of RNA or DNA molecules of a specific type and/or sequence in a sample of interest. We have developed an assay system to accurately quantitate PCR products that util...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Martin CS,Butler L,Bronstein I

    更新日期:1995-05-01 00:00:00

  • Beta-lactamase: an ideal reporter system for monitoring gene expression in live eukaryotic cells.

    abstract::To gain insightful information about the mechanisms through which genes are activated and repressed requires gene reporter systems that are sensitive, robust, and cost-effective. Although numerous reporter gene technologies are commercially available, none are as sophisticated and user-friendly as beta-lactamase (BLA)...

    journal_title:BioTechniques

    pub_type: 杂志文章,评审

    doi:10.2144/000112292

    authors: Qureshi SA

    更新日期:2007-01-01 00:00:00

  • Modification of an automated liquid-handling system for reagent-jet, nanoliter-level dispensing.

    abstract::Reducing the scale of biochemical reactions is becoming commonplace. Examples include the screening of large libraries of chemical compounds or gene sequences. These applications demand the ability to transfer sub-microliter volumes of fluid. To this end, we have modified a Hamilton MICROLAB 2200 with high-speed solen...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/01304rr05

    authors: Hicks JS,Harker BW,Beattie KL,Doktycz MJ

    更新日期:2001-04-01 00:00:00

  • Enhanced assessment of contractile dynamics in Drosophila hearts.

    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

    pub_type: 杂志文章

    doi:10.2144/000114255

    authors: Cammarato A,Ocorr S,Ocorr K

    更新日期:2015-02-01 00:00:00

  • The CTAB-DNA precipitation method: a common mini-scale preparation of template DNA from phagemids, phages or plasmids suitable for sequencing.

    abstract::This report describes a common method of obtaining template DNA from phagemids, phages and plasmids. The strategy is based on the use of the cationic detergent cetyl-trimethylammonium bromide (CTAB) for DNA precipitation. By avoiding phase separation, many manipulation steps are reduced. A time-saving modification to ...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Del Sal G,Manfioletti G,Schneider C

    更新日期:1989-05-01 00:00:00

  • Localization of trinucleotide repeat sequences in myotonic dystrophy cells using a single fluorochrome-labeled PNA probe.

    abstract::A labeled peptide nucleic acid (PNA) antisense probe was used to study the spatial distribution of triplet repeats (CTG) in human myotonic dystrophy (DM) cells by high-resolution fluorescence in situ hybridization (FISH). It was found that transcripts containing triplet repeats were present as a number of discrete foc...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/98243rr02

    authors: Taneja KL

    更新日期:1998-03-01 00:00:00

  • Antibiotic-free bacterial strain selection using antisense peptide nucleic acid.

    abstract::Antibiotics are widely useful in medicine, agriculture, and industrial fermentations. However, increasing problems with resistant strains call for restrained use and alternative strategies. Antisense peptide nucleic acids (PNAs) show potent bactericidal effects when targeted against the essential Escherichia coli acpP...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Dryselius R,Nekhotiaeva N,Nielsen PE,Good L

    更新日期:2003-11-01 00:00:00