Nonradioactive detection of telomerase activity using the telomeric repeat amplification protocol.

Abstract:

:The telomeric repeat amplification protocol (TRAP) is a two-step process for analyzing telomerase activity in cell or tissue extracts. Recent modifications of this sensitive assay include elimination of radioactivity by using a fluorescently labeled primer instead of a radiolabeled primer. In addition, the TRAP assay has been modified for real-time, quantitative PCR analysis. Here, we describe cost-effective procedures for detection of telomerase activity using a fluorescent-based assay as well as by using real-time PCR. These modified TRAP assays can be accomplished within 4 h (from lysis of samples to analysis of telomerase products).

journal_name

Nat Protoc

journal_title

Nature protocols

authors

Herbert BS,Hochreiter AE,Wright WE,Shay JW

doi

10.1038/nprot.2006.239

subject

Has Abstract

pub_date

2006-01-01 00:00:00

pages

1583-90

issue

3

eissn

1754-2189

issn

1750-2799

pii

nprot.2006.239

journal_volume

1

pub_type

杂志文章
  • Synthesis of alkyl- and aryl-amino-substituted anthraquinone derivatives by microwave-assisted copper(0)-catalyzed Ullmann coupling reactions.

    abstract::This protocol describes the efficient, generally applicable Ullmann coupling reaction of bromaminic acid with alkyl- or aryl-amines in phosphate buffer under microwave irradiation using elemental copper as a catalyst. The reaction leads to a number of biologically active compounds. As a prototypical example, the synth...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2010.63

    authors: Baqi Y,Müller CE

    更新日期:2010-05-01 00:00:00

  • A plate-based electrochromic approach for the high-throughput detection of electrochemically active bacteria.

    abstract::Electrochemically active bacteria (EAB) have the ability to transfer electrons to electron acceptors located outside the cell, and they are widely present in diverse environments. In spite of their important roles in geochemical cycles, environmental remediation and electricity generation, so far, only a limited numbe...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2013.173

    authors: Yuan SJ,Li WW,Cheng YY,He H,Chen JJ,Tong ZH,Lin ZQ,Zhang F,Sheng GP,Yu HQ

    更新日期:2014-01-01 00:00:00

  • Implementation of a 4Pi-SMS super-resolution microscope.

    abstract::The development of single-molecule switching (SMS) fluorescence microscopy (also called single-molecule localization microscopy) over the last decade has enabled researchers to image cell biological structures at unprecedented resolution. Using two opposing objectives in a so-called 4Pi geometry doubles the available ...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/s41596-020-00428-7

    authors: Wang J,Allgeyer ES,Sirinakis G,Zhang Y,Hu K,Lessard MD,Li Y,Diekmann R,Phillips MA,Dobbie IM,Ries J,Booth MJ,Bewersdorf J

    更新日期:2020-12-16 00:00:00

  • The power of multiplexed functional analysis of genetic variants.

    abstract::New technologies have recently enabled saturation mutagenesis and functional analysis of nearly all possible variants of regulatory elements or proteins of interest in single experiments. Here we discuss the past, present, and future of such multiplexed (functional) assays for variant effects (MAVEs). MAVEs provide de...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2016.135

    authors: Gasperini M,Starita L,Shendure J

    更新日期:2016-10-01 00:00:00

  • A primary CD4(+) T cell model of HIV-1 latency established after activation through the T cell receptor and subsequent return to quiescence.

    abstract::A mechanistic understanding of HIV-1 latency depends on a model system that recapitulates the in vivo condition of latently infected, resting CD4(+) T lymphocytes. Latency seems to be established after activated CD4(+) T cells, the principal targets of HIV-1 infection, become productively infected and survive long eno...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2014.188

    authors: Kim M,Hosmane NN,Bullen CK,Capoferri A,Yang HC,Siliciano JD,Siliciano RF

    更新日期:2014-12-01 00:00:00

  • Study of in vivo catheter biofilm infections using pediatric central venous catheter implanted in rat.

    abstract::Venous access catheters used in clinics are prone to biofilm contamination, contributing to chronic and nosocomial infections. Although several animal models for studying device-associated biofilms were previously described, only a few detailed protocols are currently available. Here we provide a protocol using totall...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2016.033

    authors: Chauhan A,Ghigo JM,Beloin C

    更新日期:2016-03-01 00:00:00

  • Synthesis of complex head-to-side-chain cyclodepsipeptides.

    abstract::Cyclodepsipeptides are cyclic peptides in which at least one amide link on the backbone is replaced with an ester link. These natural products present a high structural diversity that corresponds to a broad range of biological activities. Therefore, they are very promising pharmaceutical candidates. Most of the cyclod...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2016.116

    authors: Pelay-Gimeno M,Albericio F,Tulla-Puche J

    更新日期:2016-10-01 00:00:00

  • Comprehensive structural glycomic characterization of the glycocalyxes of cells and tissues.

    abstract::The glycocalyx comprises glycosylated proteins and lipids and fcorms the outermost layer of cells. It is involved in fundamental inter- and intracellular processes, including non-self-cell and self-cell recognition, cell signaling, cellular structure maintenance, and immune protection. Characterization of the glycocal...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/s41596-020-0350-4

    authors: Li Q,Xie Y,Wong M,Barboza M,Lebrilla CB

    更新日期:2020-08-01 00:00:00

  • In vivo two-photon imaging of sensory-evoked dendritic calcium signals in cortical neurons.

    abstract::Neurons in cortical sensory regions receive modality-specific information through synapses that are located on their dendrites. Recently, the use of two-photon microscopy combined with whole-cell recordings has helped to identify visually evoked dendritic calcium signals in mouse visual cortical neurons in vivo. The c...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2010.169

    authors: Jia H,Rochefort NL,Chen X,Konnerth A

    更新日期:2011-01-01 00:00:00

  • Development of a highly reproducible three-dimensional organotypic model of the oral mucosa.

    abstract::In this report we describe the development of a standardized three-dimensional (3D) system of the human oral mucosa based on an immortalized human oral keratinocyte cell line (OKF6/TERT-2). The procedure takes approximately 2-3 weeks to complete and includes three main stages: preparation of collagen-embedded fibrobla...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2006.323

    authors: Dongari-Bagtzoglou A,Kashleva H

    更新日期:2006-01-01 00:00:00

  • In vitro 'sexual' evolution through the PCR-based staggered extension process (StEP).

    abstract::This protocol describes a directed evolution method for in vitro mutagenesis and recombination of polynucleotide sequences. The staggered extension process (StEP) is essentially a modified PCR that uses highly abbreviated annealing and extension steps to generate staggered DNA fragments and promote crossover events al...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2006.309

    authors: Zhao H,Zha W

    更新日期:2006-01-01 00:00:00

  • Structure determination protocol for transmembrane domain oligomers.

    abstract::The transmembrane (TM) anchors of cell surface proteins have been one of the 'blind spots' in structural biology because they are generally very hydrophobic, sometimes dynamic, and thus difficult targets for structural characterization. A plethora of examples show these membrane anchors are not merely anchors but can ...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/s41596-019-0188-9

    authors: Fu Q,Piai A,Chen W,Xia K,Chou JJ

    更新日期:2019-08-01 00:00:00

  • TARDIS, a targeted RNA directional sequencing method for rare RNA discovery.

    abstract::High-throughput transcriptional analysis has unveiled a myriad of novel RNAs. However, technical constraints in RNA sequencing library preparation and platform performance hamper the identification of rare transcripts contained within the RNA repertoire. Herein we present targeted-RNA directional sequencing (TARDIS), ...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2015.120

    authors: Portal MM,Pavet V,Erb C,Gronemeyer H

    更新日期:2015-12-01 00:00:00

  • In-gel stable isotope labeling for relative quantification using mass spectrometry.

    abstract::Although differences in protein staining intensity can often be visualized by difference gel electrophoresis, abundant proteins can obscure less abundant proteins, and quantification of post-translational modifications is difficult. We present a protocol for quantifying changes in the abundance of a specific protein o...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2006.7

    authors: Asara JM,Zhang X,Zheng B,Maroney LA,Christofk HR,Wu N,Cantley LC

    更新日期:2006-01-01 00:00:00

  • Real-time single-molecule coimmunoprecipitation of weak protein-protein interactions.

    abstract::Coimmunoprecipitation (co-IP) analysis is a useful method for studying protein-protein interactions. It currently involves electrophoresis and western blotting, which are not optimized for detecting weak and transient interactions. In this protocol we describe an advanced version of co-IP analysis that uses real-time,...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2013.116

    authors: Lee HW,Ryu JY,Yoo J,Choi B,Kim K,Yoon TY

    更新日期:2013-10-01 00:00:00

  • Production of a site- and phosphorylation state-specific antibody.

    abstract::Protein phosphorylation plays important roles in various aspects of cellular events. Visualization of site-specific phosphorylation in cells is of great importance not only to analyze spatial and temporal distribution but also to investigate biological function. Now, site- and phosphorylation state-specific antibodies...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2007.374

    authors: Goto H,Inagaki M

    更新日期:2007-01-01 00:00:00

  • Cognitive analysis of metabolomics data for systems biology.

    abstract::Cognitive computing is revolutionizing the way big data are processed and integrated, with artificial intelligence (AI) natural language processing (NLP) platforms helping researchers to efficiently search and digest the vast scientific literature. Most available platforms have been developed for biomedical researcher...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/s41596-020-00455-4

    authors: Majumder EL,Billings EM,Benton HP,Martin RL,Palermo A,Guijas C,Rinschen MM,Domingo-Almenara X,Montenegro-Burke JR,Tagtow BA,Plumb RS,Siuzdak G

    更新日期:2021-01-22 00:00:00

  • High spatial resolution imaging of biological tissues using nanospray desorption electrospray ionization mass spectrometry.

    abstract::Mass spectrometry imaging (MSI) enables label-free spatial mapping of hundreds of biomolecules in tissue sections. This capability provides valuable information on tissue heterogeneity that is difficult to obtain using population-averaged assays. Despite substantial developments in both instrumentation and methodology...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/s41596-019-0237-4

    authors: Yin R,Burnum-Johnson KE,Sun X,Dey SK,Laskin J

    更新日期:2019-12-01 00:00:00

  • Transient expression and analysis of fluorescent reporter proteins in plant pollen tubes.

    abstract::The pollen tube is an excellent single-cell model system for studying cellular processes in plant cell biology. This protocol describes a detailed step-by-step procedure with optimized conditions for introducing various fluorescent reporter proteins into lily, tobacco and Arabidopsis pollen grains by means of biolisti...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2011.309

    authors: Wang H,Jiang L

    更新日期:2011-04-01 00:00:00

  • Fluid-percussion-induced traumatic brain injury model in rats.

    abstract::Traumatic brain injury (TBI) is a major cause of mortality and morbidity. Various attempts have been made to replicate clinical TBI using animal models. The fluid-percussion model (FP) is one of the oldest and most commonly used models of experimentally induced TBI. Both central (CFP) and lateral (LFP) variations of t...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2010.112

    authors: Kabadi SV,Hilton GD,Stoica BA,Zapple DN,Faden AI

    更新日期:2010-09-01 00:00:00

  • Mapping 3D genome architecture through in situ DNase Hi-C.

    abstract::With the advent of massively parallel sequencing, considerable work has gone into adapting chromosome conformation capture (3C) techniques to study chromosomal architecture at a genome-wide scale. We recently demonstrated that the inactive murine X chromosome adopts a bipartite structure using a novel 3C protocol, ter...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2016.126

    authors: Ramani V,Cusanovich DA,Hause RJ,Ma W,Qiu R,Deng X,Blau CA,Disteche CM,Noble WS,Shendure J,Duan Z

    更新日期:2016-11-01 00:00:00

  • Oxidative bisulfite sequencing of 5-methylcytosine and 5-hydroxymethylcytosine.

    abstract::To uncover the function of and interplay between the mammalian cytosine modifications 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC), new techniques and advances in current technology are needed. To this end, we have developed oxidative bisulfite sequencing (oxBS-seq), which can quantitatively locate 5mC an...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2013.115

    authors: Booth MJ,Ost TW,Beraldi D,Bell NM,Branco MR,Reik W,Balasubramanian S

    更新日期:2013-10-01 00:00:00

  • Coupling surface plasmon resonance to mass spectrometry to discover novel protein-protein interactions.

    abstract::The elucidation of protein-protein interaction networks is a crucial task in the postgenomic era. In this protocol, we describe our approach to discover protein-protein interactions using the surface plasmon resonance technique coupled to mass spectrometry (MS). A peptide or a protein is immobilized on a sensor chip a...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2009.84

    authors: Madeira A,Ohman E,Nilsson A,Sjögren B,Andrén PE,Svenningsson P

    更新日期:2009-01-01 00:00:00

  • Pulsed-field gel electrophoresis.

    abstract::This protocol describes pulsed-field gel electrophoresis (PFGE), a method developed for separation of large DNA molecules. Whereas standard DNA gel electrophoresis commonly resolves fragments up to approximately 50 kb in size, PFGE fractionates DNA molecules up to 10 Mb. The mechanism driving these separations exploit...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2007.94

    authors: Herschleb J,Ananiev G,Schwartz DC

    更新日期:2007-01-01 00:00:00

  • Femtosecond optical transfection of individual mammalian cells.

    abstract::Laser-mediated gene transfection into mammalian cells has recently emerged as a powerful alternative to more traditional transfection techniques. In particular, the use of a femtosecond-pulsed laser operating in the near-infrared (NIR) region has been proven to provide single-cell selectivity, localized delivery, low ...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2013.071

    authors: Antkowiak M,Torres-Mapa ML,Stevenson DJ,Dholakia K,Gunn-Moore FJ

    更新日期:2013-06-01 00:00:00

  • Measuring bone blood supply in mice using fluorescent microspheres.

    abstract::Fluorescent microspheres are commonly used to assess bone blood supply in large animals, but the technique is not widely used in smaller mammals, as traditional methods such as reference blood sampling, ventilation and catheterization are not easily applied. This protocol describes a viable alternative for measuring b...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2009.190

    authors: Serrat MA

    更新日期:2009-01-01 00:00:00

  • Derivation and maintenance of human embryonic stem cells from poor-quality in vitro fertilization embryos.

    abstract::Human embryonic stem (hES) cells are self-renewing, pluripotent cells that are valuable research tools and hold promise for use in regenerative medicine. Most hES cell lines are derived from cryopreserved human embryos that were created during in vitro fertilization (IVF) and are in excess of clinical need. Embryos th...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2008.60

    authors: Lerou PH,Yabuuchi A,Huo H,Miller JD,Boyer LF,Schlaeger TM,Daley GQ

    更新日期:2008-01-01 00:00:00

  • A mass spectrometry workflow for measuring protein turnover rates in vivo.

    abstract::Proteins are continually produced and degraded, to avoid the accumulation of old or damaged molecules and to maintain the efficiency of physiological processes. Despite its importance, protein turnover has been difficult to measure in vivo. Previous approaches to evaluating turnover in vivo have required custom labeli...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/s41596-019-0222-y

    authors: Alevra M,Mandad S,Ischebeck T,Urlaub H,Rizzoli SO,Fornasiero EF

    更新日期:2019-12-01 00:00:00

  • Agrobacterium-mediated transformation of rice using immature embryos or calli induced from mature seed.

    abstract::Here, we provide comprehensive, highly efficient protocols for Agrobacterium tumefaciens-mediated transformation of a wide range of rice genotypes. Methods that use either immature embryos (japonica and indica rice) or calli (japonica cultivars and the indica cultivar, Kasalath) as a starting material for inoculation ...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2008.46

    authors: Hiei Y,Komari T

    更新日期:2008-01-01 00:00:00

  • Advanced CUBIC protocols for whole-brain and whole-body clearing and imaging.

    abstract::Here we describe a protocol for advanced CUBIC (Clear, Unobstructed Brain/Body Imaging Cocktails and Computational analysis). The CUBIC protocol enables simple and efficient organ clearing, rapid imaging by light-sheet microscopy and quantitative imaging analysis of multiple samples. The organ or body is cleared by im...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2015.085

    authors: Susaki EA,Tainaka K,Perrin D,Yukinaga H,Kuno A,Ueda HR

    更新日期:2015-11-01 00:00:00