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; however, it has been adjusted because of the large quantities of carbohydrate and nucleases found in Dictyostelium cells. After cell lysis and phenol:chloroform extraction, the resulting high-quality RNA isolated with the described protocol allows the molecular genetic analysis of wild-type and genetically modified cells. The purified RNA can be used for analyses such as northern blotting, RT-PCR and microarrays. This procedure requires approximately 2 h to complete.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Pilcher KE,Gaudet P,Fey P,Kowal AS,Chisholm RLdoi
10.1038/nprot.2007.191subject
Has Abstractpub_date
2007-01-01 00:00:00pages
1329-32issue
6eissn
1754-2189issn
1750-2799pii
nprot.2007.191journal_volume
2pub_type
杂志文章相关文献
Nature Protocols文献大全abstract：:Realizing the potential of human embryonic stem cells (hESCs) in research and commercial applications requires generic protocols for culture, expansion and genetic modification that function between multiple lines. Here we describe a feeder-free hESC culture protocol that was tested in 13 independent hESC lines derive...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2008.140
更新日期：2008-01-01 00:00:00
abstract：:The human stomach contains two primary domains: the corpus, which contains the fundic epithelium, and the antrum. Each of these domains has distinct cell types and functions, and therefore each presents with unique disease pathologies. Here, we detail two protocols to differentiate human pluripotent stem cells (hPSCs)...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-018-0080-z
更新日期：2019-01-01 00:00:00
abstract：:Establishing a small animal model that accurately recapitulates hepatotropic pathogens, including hepatitis C virus (HCV) infection and immunopathogenesis, is essential for the study of hepatitis virus-induced liver disease and for therapeutics development. This protocol describes our recently developed humanized mous...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2012.083
更新日期：2012-09-01 00:00:00
abstract：:Over the past 20 years, the starlet sea anemone, Nematostella vectensis, a small estuarine animal, has emerged as a powerful model system for field and laboratory studies of development, evolution, genomics, molecular biology and toxicology. Here we describe how to collect Nematostella, culture it through its entire s...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2013.044
更新日期：2013-05-01 00:00:00
abstract：:Apoptosis plays a pivotal role in the regulation of cell turnover, and a defect or an excess of apoptosis has been implicated in several human diseases. Apoptosis is activated from an extracellular death signal, or from an internal pathway starting from the endoplasmatic reticulum or the mitochondria. To investigate t...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.397
更新日期：2007-01-01 00:00:00
abstract：:The unprecedented increase in the throughput of DNA sequencing driven by next-generation technologies now allows efficient analysis of the complete protein-coding regions of genomes (exomes) for multiple samples in a single sequencing run. However, sample preparation and targeted enrichment of multiple samples has bec...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2011.396
更新日期：2011-11-03 00:00:00
abstract：:We describe a step-by-step protocol for measuring the stable products of the nitric oxide (NO) pathway: nitrite, nitrite plus nitrate and nitrate. This described protocol is easy to apply and is about 50 times more sensitive than the commonly used Griess reaction or commercially available assay kits based on the Gries...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.341
更新日期：2006-01-01 00:00:00
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
更新日期：2014-01-01 00:00:00
abstract：:The small ubiquitin-like modifiers (SUMOs) are posttranslationally conjugated to eukaryotic cellular proteins with generally unpredictable consequences. SUMO substrates are found in many cellular systems, and functional analysis has revealed that substrate SUMOylation often has an important role in their regulation. H...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2009.128
更新日期：2009-01-01 00:00:00
abstract：:The Golgi apparatus undergoes extensive disassembly during mitosis and reassembly in post-mitotic daughter cells. This process has been mimicked in vitro by treating Golgi membranes with mitotic and interphase cytosol. To determine the minimal machinery that controls the morphological change, we have developed a defin...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2010.38
更新日期：2010-04-01 00:00:00
abstract：:This protocol is an extension to: Nat. Protoc. 6, 870-895 (2014); doi:10.1038/nprot.2011.328; published online 02 June 2011Cycling cells duplicate their DNA content during S phase, following a defined program called replication timing (RT). Early- and late-replicating regions differ in terms of mutation rates, transcr...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2017.148
更新日期：2018-05-01 00:00:00
abstract：:The Strep-tag II is an eight-residue minimal peptide sequence (Trp-Ser-His-Pro-Gln-Phe-Glu-Lys) that exhibits intrinsic affinity toward streptavidin and can be fused to recombinant proteins in various fashions. We describe a protocol that enables quick and mild purification of corresponding Strep-tag II fusion protein...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.209
更新日期：2007-01-01 00:00:00
abstract：:This protocol describes a method for direct labeling and detection of small RNAs present in total RNA by splinted ligation. The assay uses a small RNA-specific bridge oligonucleotide to form base pairs with the small RNA and a 5'-end-radiolabeled ligation oligonucleotide. The captured small RNA is directly labeled by ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.530
更新日期：2008-01-01 00:00:00
abstract：:Fast, high-resolution mapping of heterogeneous interfaces with a wide elastic modulus range is a major goal of atomic force microscopy (AFM). This goal becomes more challenging when the nanomechanical mapping involves biomolecules in their native environment. Over the years, several AFM-based methods have been develop...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-018-0070-1
更新日期：2018-12-01 00:00:00
abstract：:RNA sequencing (RNA-seq) has been rapidly adopted for the profiling of transcriptomes in many areas of biology, including studies into gene regulation, development and disease. Of particular interest is the discovery of differentially expressed genes across different conditions (e.g., tissues, perturbations) while opt...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2013.099
更新日期：2013-09-01 00:00:00
abstract：:The study of angiogenic endothelial cells (ECs) has in recent years greatly stimulated multiple fields of vascular biology research. A number of cellular models and numerous complex developmental, manipulatory and tumor animal models have been developed to study angiogenesis in vitro and in vivo. To connect the versat...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2009.96
更新日期：2009-01-01 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：:The membrane-permeant fluorogenic biarsenicals FlAsH-EDT(2) and ReAsH-EDT(2) can be prepared in good yields by a straightforward two-step procedure from the inexpensive precursor dyes fluorescein and resorufin, respectively. Handling of toxic reagents such as arsenic trichloride is minimized so the synthesis can be ca...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2008.144
更新日期：2008-01-01 00:00:00
abstract：:Association studies can focus on candidate gene(s), a particular genomic region, or adopt a genome-wide association approach, each of which has implications for marker selection. The strategy for marker selection will affect the statistical power of the study to detect a disease association and is a crucial element of...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2009.38
更新日期：2009-01-01 00:00:00
abstract：:The CRISPR-Cas9 genome-editing tool and the availability of whole-genome sequences from plant species have revolutionized our ability to introduce targeted mutations into important crop plants, both to explore genetic changes and to introduce new functionalities. Here, we describe protocols adapting the CRISPR-Cas9 sy...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-018-0067-9
更新日期：2018-12-01 00:00:00
abstract：:Determining enantiomeric excess (e.e.) in chiral compounds is key to development of chiral catalyst auxiliaries and chiral drugs. Here we describe a sensitive and robust fluorescence-based assay for determining e.e. in mixtures of enantiomers of 1,2- and 1,3-diols, chiral amines, amino alcohols, and amino-acid esters....
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-020-0329-1
更新日期：2020-07-01 00:00:00
abstract：:This protocol and the accompanying software program called LEVER (lineage editing and validation) enable quantitative automated analysis of phase-contrast time-lapse images of cultured neural stem cells. Images are captured at 5-min intervals over a period of 5-15 d as the cells proliferate and differentiate. LEVER au...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2011.422
更新日期：2011-11-17 00:00:00
abstract：:This protocol describes a culture system in which inner-ear sensory tissue is produced from mouse embryonic stem (ES) cells under chemically defined conditions. This model is amenable to basic and translational investigations into inner ear biology and regeneration. In this protocol, mouse ES cells are aggregated in 9...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2014.100
更新日期：2014-01-01 00:00:00
abstract：:The plant cell wall is a chemically complex structure composed mostly of polysaccharides. Detailed analyses of these cell wall polysaccharides are essential for our understanding of plant development and for our use of plant biomass (largely wall material) in the food, agriculture, fabric, timber, biofuel and biocompo...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2012.081
更新日期：2012-09-01 00:00:00
abstract：:ErmineJ is software for the analysis of functionally interesting patterns in large gene lists drawn from gene expression profiling data or other high-throughput genomics studies. It can be used by biologists with no bioinformatics background to conduct sophisticated analyses of gene sets with multiple methods. It allo...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2010.78
更新日期：2010-06-01 00:00:00
abstract：:Transitivity Clustering is a method for the partitioning of biological data into groups of similar objects, such as genes, for instance. It provides integrated access to various functions addressing each step of a typical cluster analysis. To facilitate this, Transitivity Clustering is accessible online and offers thr...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2010.197
更新日期：2011-03-01 00:00:00
abstract：:Lentiviral vectors derived from the human immunodeficiency type 1 virus (HIV-1 LV) are among the finest tools available today for the genetic modification of human monocyte-derived dendritic cells (MDDCs). However, this process is largely inefficient because MDDCs show a strong resistance to HIV-1 transduction. Here w...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2011.327
更新日期：2011-06-01 00:00:00
abstract：:An optimized procedure for the efficient preparation of 2-heptyl-3-hydroxy-4(1H)-quinolone (Pseudomonas quinolone signal or PQS) and a diverse range of structurally related 2-alkyl-4-quinolones with biological activity is presented. The two-step synthesis begins with the formation of α-chloro ketones by the coupling o...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2012.054
更新日期：2012-05-24 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：: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
更新日期：2006-01-01 00:00:00
