Abstract：
:Telomeres have emerged as crucial cellular elements in aging and various diseases including cancer. To measure the average length of telomere repeats in cells, we describe our protocols that use fluorescent in situ hybridization (FISH) with labeled peptide nucleic acid (PNA) probes specific for telomere repeats in combination with fluorescence measurements by flow cytometry (flow FISH). Flow FISH analysis can be performed using commercially available flow cytometers, and has the unique advantage over other methods for measuring telomere length of providing multi-parameter information on the length of telomere repeats in thousands of individual cells. The accuracy and reproducibility of the measurements is augmented by the automation of most pipetting (aspiration and dispensing) steps, and by including an internal standard (control cells) with a known telomere length in every tube. The basic protocol for the analysis of nucleated blood cells from 22 different individuals takes about 12 h spread over 2-3 days.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Baerlocher GM,Vulto I,de Jong G,Lansdorp PMdoi
10.1038/nprot.2006.263subject
Has Abstractpub_date
2006-01-01 00:00:00pages
2365-76issue
5eissn
1754-2189issn
1750-2799pii
nprot.2006.263journal_volume
1pub_type
杂志文章相关文献
Nature Protocols文献大全abstract：:Most currently available colorectal cancer (CRC) mouse models are not suitable for studying progression toward the metastatic stage. Recently, establishment of tumor organoid lines, either from murine CRC models or patients, and the possibility of engineering them with genome-editing technologies, have provided a larg...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2017.137
更新日期：2018-02-01 00:00:00
abstract：:Here we present a protocol for labeling aliphatic carboxylic acids with the positron-emitting radionuclide 11C (t(1/2) = 20.4 min) at the carboxyl position using [11C]carbon monoxide via photoinitiated free radical-mediated carbonylation. A solution of an alkyl iodide in a homogenous binary organic solvent-water mixtu...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.112
更新日期：2006-01-01 00:00:00
abstract：:Hepatic stellate cells (HSCs) have been identified as the main fibrogenic cell type in the liver. Hence, efforts to understand hepatic fibrogenesis and to develop treatment strategies have focused on this cell type. HSC isolation, originally developed in rats, has subsequently been adapted to mice, thus allowing the s...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2015.017
更新日期：2015-02-01 00:00:00
abstract：:Genome editing harnesses programmable nucleases to cut and paste genetic information in a targeted manner in living cells and organisms. Here, I review the development of programmable nucleases, including zinc finger nucleases (ZFNs), TAL (transcription-activator-like) effector nucleases (TALENs) and CRISPR (cluster o...
journal_title：Nature protocols
pub_type： 杂志文章,评审
doi：10.1038/nprot.2016.104
更新日期：2016-09-01 00:00:00
abstract：:Carbohydrate microarrays have received considerable attention as an advanced technology for the rapid analysis of carbohydrate-protein interactions. This protocol provides detailed procedures for the preparation of carbohydrate microarrays by immobilizing hydrazide-conjugated carbohydrates on epoxide-derivatized glass...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.373
更新日期：2007-01-01 00:00:00
abstract：:In this report, we describe a two-step protocol for labeling of an affinity-purified antibody to biotin with horseradish peroxidase (HRP) using cyanuric chloride (CC) as a bridge. The enzyme was first modified with CC, and following chromatography on a PD-10 column, the activated HRP was incubated with the antibody to...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2009.6
更新日期：2009-01-01 00:00:00
abstract：:Reactive oxygen species (ROS) are continuously produced in the cell as a consequence of aerobic metabolism, and are controlled by several antioxidant mechanisms. An accurate measurement of ROS is essential to evaluate the redox status of the cell, or the effects of molecules with the pro-oxidant or antioxidant activit...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2009.189
更新日期：2009-01-01 00:00:00
abstract：:Here we present a protocol for the extraction of RNA from Dictyostelium discoideum. Dictyostelium is a social amoeba that undergoes a basic developmental program, and therefore analysis of RNA levels over a time course is a commonly used technique. This procedure is similar to other guanidine thiocyanate-based methods...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.191
更新日期：2007-01-01 00:00:00
abstract：:This protocol describes the synthesis of oligo(poly(ethylene glycol) fumarate) (OPF; 1-35 kDa; a polymer useful for tissue engineering applications) by a one-pot reaction of poly(ethylene glycol) (PEG) and fumaryl chloride. The procedure involves three parts: dichloromethane and PEG are first dried; the reaction step ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2012.055
更新日期：2012-05-31 00:00:00
abstract：:One of the challenges for modern neuroscience is to understand the rules of concerted neuronal function in vivo. This question can be addressed using noninvasive high-resolution imaging techniques like two-photon microscopy. This protocol describes a versatile approach for in vivo two-photon calcium imaging of neural ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.58
更新日期：2006-01-01 00:00:00
abstract：:Precise identification of sites of RNA modification is key to studying the functional role of such modifications in the regulation of gene expression and for elucidating relevance to diverse physiological processes. tRNA reduction and cleavage sequencing (TRAC-Seq) is a chemically based approach for the unbiased globa...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-019-0226-7
更新日期：2019-11-01 00:00:00
abstract：:Competition chromatin immunoprecipitation (competition ChIP) enables experimenters to measure protein-DNA dynamics at a single locus or across the entire genome, depending on the detection method. Competition ChIP relies on a cell containing two copies of a single DNA-associated factor, with each copy of the factor di...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2013.077
更新日期：2013-01-01 00:00:00
abstract：:This protocol describes a method for directing the expression of genes of interest into postmitotic neocortical neurons in vivo. Microinjection of a DNA plasmid-amphiphilic molecule mix into the neocortex followed by delivery of an ad hoc electric pulse protocol during the first few days of life in mice allows rapid, ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2015.001
更新日期：2015-01-01 00:00:00
abstract：:In situ mRNA hybridization is one of the most powerful techniques for analyzing patterns of gene expression. However, its usefulness is limited in complex plant tissues by the need to fix, embed and section samples before localizing the desired mRNA. Here we present a robust whole-mount in situ hybridization method th...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2014.162
更新日期：2014-10-01 00:00:00
abstract：:Here we describe a protocol for the production of frozen competent yeast cells that can be transformed with high efficiency using the lithium acetate/single-stranded carrier DNA/PEG method. This protocol allows the production of highly competent yeast cells that can be frozen and used at a later date and is especially...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.17
更新日期：2007-01-01 00:00:00
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
更新日期：2011-01-01 00:00:00
abstract：:Conventional gene expression studies analyze multiple cells simultaneously or single cells, for which the exact in vivo or in situ position is unknown. Although cellular heterogeneity can be discerned when analyzing single cells, any spatially defined attributes that underpin the heterogeneous nature of the cells cann...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2017.003
更新日期：2017-03-01 00:00:00
abstract：:Using siRNAs to genetically manipulate immune cells is important to both basic immunological studies and therapeutic applications. However, siRNA delivery is challenging because primary immune cells are often sensitive to the delivery materials and generate immune responses. We have recently developed an amphiphilic d...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-020-00418-9
更新日期：2021-01-01 00:00:00
abstract：:Carbon-based dots (CDs) and their functionalized (nano)composites have recently attracted attention due to their seemingly easy preparation and numerous potential applications, ranging from those in the biomedical field (i.e., imaging and drug delivery) to those in (opto)electronics (i.e., solar cells and LEDs). This ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-019-0207-x
更新日期：2019-10-01 00:00:00
abstract：:The bimolecular fluorescence complementation (BiFC) assay is a powerful tool for visualizing and identifying protein interactions in living cells. This assay is based on the principle of protein-fragment complementation, using two nonfluorescent fragments derived from fluorescent proteins. When two fragments are broug...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2008.16
更新日期：2008-01-01 00:00:00
abstract：:Chromatin and transcriptional processes are among the most intensively studied fields of biology today. The introduction of chromatin immunoprecipitations (ChIP) represents a major advancement in this area. This powerful method allows researchers to probe specific protein-DNA interactions in vivo and to estimate the d...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.27
更新日期：2006-01-01 00:00:00
abstract：:Studies on colonic cells in the lamina propria (LP) of mice are important for understanding the cellular and immune responses in the gut, especially in inflammatory bowel diseases (such as morbus crohn and colitis ulcerosa). This protocol details a method to isolate LP cells and characterize freshly isolated cells by ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.315
更新日期：2007-01-01 00:00:00
abstract：:The small ubiquitin-like modifier (SUMO) is a protein modifier that is post-translationally coupled to thousands of lysines in more than a thousand proteins. An understanding of which lysines are modified by SUMO is critical in unraveling its function as a master regulator of all nuclear processes, as well as its invo...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2016.082
更新日期：2016-09-01 00:00:00
abstract：:Targeted nucleases, including zinc-finger nucleases (ZFNs), transcription activator-like (TAL) effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9), have provided researchers with the ability to manipulate nearly any genomic sequence in hu...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2015.117
更新日期：2015-11-01 00:00:00
abstract：:Multiplex-fluorescence in situ hybridization (M-FISH) was initially developed to stain human chromosomes--the 22 autosomes and X and Y sex chromosomes--with uniquely distinctive colors to facilitate karyotyping. The characteristic spectral signatures of all different combinations of fluorochromes are determined by mul...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.160
更新日期：2006-01-01 00:00:00
abstract：:The ability to isolate and analyze rare circulating tumor cells (CTCs) has the potential to further our understanding of cancer metastasis and enhance the care of cancer patients. In this protocol, we describe the procedure for isolating rare CTCs from blood samples by using tumor antigen-independent microfluidic CTC-...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2014.044
更新日期：2014-03-01 00:00:00
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
更新日期：2008-01-01 00:00:00
abstract：:Many techniques have been developed for studying inducible gene expression, but all of them are multicomponent systems consisting of cis-acting elements at the DNA or RNA level, trans-acting regulator proteins and/or small molecules as inducers. RNA thermometers are the only known single-component regulators of gene e...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2009.112
更新日期：2009-01-01 00:00:00
abstract：:Freestanding plasmonic nanoparticle (NP) superlattice sheets are novel 2D nanomaterials with tailorable properties that enable their use for broad applications in sensing, anticounterfeit measures, ionic gating, nanophotonics and flat lenses. We recently developed a robust, yet general, two-step drying-mediated approa...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-019-0200-4
更新日期：2019-09-01 00:00:00
abstract：:In the past few decades, marine natural products bioprospecting has yielded a considerable number of drug candidates. Two marine natural products have recently been admitted as new drugs: Prialt (also known as ziconotide) as a potent analgesic for severe chronic pain and Yondelis (known also as trabectedin or E-743) a...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2008.182
更新日期：2008-01-01 00:00:00
