Frozen competent yeast cells that can be transformed with high efficiency using the LiAc/SS carrier DNA/PEG method.

abstract：:Cell-to-cell chemical signaling plays multiple roles in coordinating the activity of the functional elements of an organism, with these elements ranging from a three-neuron reflex circuit to the entire animal. In recent years, single-cell mass spectrometry (MS) has enabled the discovery of cell-to-cell signaling molec...

journal_title：Nature protocols

authors： Rubakhin SS,Sweedler JV

更新日期：2007-01-01 00:00:00

abstract：:Live-imaging technology has markedly advanced in the field of neural injury and axon degeneration; however, studies are still predominantly performed in in vitro settings such as cultured neuronal cells or in model organisms such as Caenorhabditis elegans in which axons lack glial wrappings. We recently developed a ne...

journal_title：Nature protocols

authors： Fang Y,Soares L,Bonini NM

更新日期：2013-04-01 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

authors： Benaglia S,Gisbert VG,Perrino AP,Amo CA,Garcia R

更新日期：2018-12-01 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

authors： Liu J,Gaj T,Yang Y,Wang N,Shui S,Kim S,Kanchiswamy CN,Kim JS,Barbas CF 3rd

更新日期：2015-11-01 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

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

abstract：:RNA production using in vivo transcription by Escherichia coli allows preparation of milligram quantities of RNA for biochemical, biophysical and structural investigations. We describe here a generic protocol for the overproduction and purification of recombinant RNA using liquid chromatography. The strategy utilizes ...

journal_title：Nature protocols

authors： Ponchon L,Beauvais G,Nonin-Lecomte S,Dardel F

更新日期：2009-01-01 00:00:00

abstract：:The analysis of highly hydrophobic proteins is still an analytical challenge. Using a recombinant gamma-aminobutyric acid A (GABAA)-receptor subunit as a model protein, we developed a gel-based proteomic approach for high MS/MS-peptide sequence coverage identification. Protein samples were separated by multi-dimension...

journal_title：Nature protocols

authors： Kang SU,Fuchs K,Sieghart W,Pollak A,Csaszar E,Lubec G

更新日期：2009-01-01 00:00:00

abstract：:This protocol describes a methodology for imaging the sequestration of infected erythrocytes of the rodent malaria parasite Plasmodium berghei in the bodies of live mice or in dissected organs, using a transgenic parasite that expresses luciferase. Real-time imaging of infected erythrocytes is performed by measuring b...

journal_title：Nature protocols

authors： Franke-Fayard B,Waters AP,Janse CJ

更新日期：2006-01-01 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

authors： Rozier F,Mirabet V,Vernoux T,Das P

更新日期：2014-10-01 00:00:00

abstract：:Mass spectrometry-based phosphoproteomic analysis is a powerful method for gaining a global, unbiased understanding of cellular signaling. Its accuracy and comprehensiveness stands or falls with the quality and choice of the applied phosphopeptide prefractionation strategy. This protocol covers a powerful but simple a...

journal_title：Nature protocols

authors： Zarei M,Sprenger A,Rackiewicz M,Dengjel J

更新日期：2016-01-01 00:00:00

abstract：:The world is on the cusp of a post-antibiotic era, but researchers and medical doctors have found a way forward-by looking back at how infections were treated before the advent of antibiotics, namely using phage therapy. Although bacteriophages (phages) continue to lack drug approval in Western medicine, an increasing...

journal_title：Nature protocols

authors： Luong T,Salabarria AC,Edwards RA,Roach DR

更新日期：2020-09-01 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

authors： Pettolino FA,Walsh C,Fincher GB,Bacic A

更新日期：2012-09-01 00:00:00

abstract：:The direct detection of drug-protein interactions in living cells is a major challenge in drug discovery research. Recently, we introduced an approach termed thermal proteome profiling (TPP), which enables the monitoring of changes in protein thermal stability across the proteome using quantitative mass spectrometry. ...

journal_title：Nature protocols

authors： Franken H,Mathieson T,Childs D,Sweetman GM,Werner T,Tögel I,Doce C,Gade S,Bantscheff M,Drewes G,Reinhard FB,Huber W,Savitski MM

更新日期：2015-10-01 00:00:00

abstract：:The mouse embryo hindbrain is a robust and adaptable model for studying sprouting angiogenesis. It permits the spatiotemporal analysis of organ vascularization in normal mice and in mouse strains with genetic mutations that result in late embryonic or perinatal lethality. Unlike postnatal models such as retinal angiog...

journal_title：Nature protocols

authors： Fantin A,Vieira JM,Plein A,Maden CH,Ruhrberg C

更新日期：2013-02-01 00:00:00

abstract：:Single-cell gene expression analysis has contributed to a better understanding of the transcriptional heterogeneity in a variety of model systems, including those used in research in developmental, cancer and stem cell biology. Nowadays, technological advances facilitate the generation of large gene expression data se...

journal_title：Nature protocols

authors： Durruthy-Durruthy R,Gottlieb A,Heller S

更新日期：2015-03-01 00:00:00

abstract：:Periodontal disease (PD) is a common dental disease associated with the interaction between dysbiotic oral microbiota and host immunity. It is a prevalent disease, resulting in loss of gingival tissue, periodontal ligament, cementum and alveolar bone. PD is a major form of tooth loss in the adult population. Experimen...

journal_title：Nature protocols

authors： Marchesan J,Girnary MS,Jing L,Miao MZ,Zhang S,Sun L,Morelli T,Schoenfisch MH,Inohara N,Offenbacher S,Jiao Y

更新日期：2018-10-01 00:00:00

abstract：:Forebrain γ-aminobutyric acid (GABA) interneurons have crucial roles in high-order brain function via modulating network activities and plasticity, and they are implicated in many psychiatric disorders. Availability of enriched functional human forebrain GABA interneurons, especially those from people affected by GABA...

journal_title：Nature protocols

authors： Liu Y,Liu H,Sauvey C,Yao L,Zarnowska ED,Zhang SC

更新日期：2013-09-01 00:00:00

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

authors： Baqi Y,Müller CE

更新日期：2010-05-01 00:00:00

abstract：:High throughput microarray transcription analyses provide us with the expression profiles for large amounts of plant genes. However, their tissue and cellular resolution is limited. Thus, for detailed functional analysis, it is still necessary to examine the expression pattern of selected candidate genes at a cellular...

journal_title：Nature protocols

authors： Hejátko J,Blilou I,Brewer PB,Friml J,Scheres B,Benková E

更新日期：2006-01-01 00:00:00

abstract：:The fabrication of 3D tissues retaining the original functions of tissues/organs in vitro is crucial for optimal tissue engineering and regenerative medicine. The fabrication of 3D tissues also contributes to the establishment of in vitro tissue/organ models for drug screening. Our laboratory has developed a fabricati...

journal_title：Nature protocols

authors： Haraguchi Y,Shimizu T,Sasagawa T,Sekine H,Sakaguchi K,Kikuchi T,Sekine W,Sekiya S,Yamato M,Umezu M,Okano T

更新日期：2012-04-05 00:00:00

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

authors： Zhao H,Zha W

更新日期：2006-01-01 00:00:00

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

authors： Fumagalli A,Suijkerbuijk SJE,Begthel H,Beerling E,Oost KC,Snippert HJ,van Rheenen J,Drost J

更新日期：2018-02-01 00:00:00

abstract：:In this protocol, we describe cryoimmunolabeling methods for the subcellular localization of proteins and certain lipids. The methods start with chemical fixation of cells and tissue in formaldehyde (FA) and/or glutaraldehyde (GA), sometimes supplemented with acrolein. Cell and tissue blocks are then immersed in 2.3 M...

journal_title：Nature protocols

authors： Slot JW,Geuze HJ

更新日期：2007-01-01 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

authors： Karabacak NM,Spuhler PS,Fachin F,Lim EJ,Pai V,Ozkumur E,Martel JM,Kojic N,Smith K,Chen PI,Yang J,Hwang H,Morgan B,Trautwein J,Barber TA,Stott SL,Maheswaran S,Kapur R,Haber DA,Toner M

更新日期：2014-03-01 00:00:00

abstract：:The accurate knowledge of receptor kinetics is crucial to our understanding of cell signal transduction in general and neural function in particular. The classical technique of probing membrane receptors on a millisecond scale involves placing a recording micropipette with a membrane patch in front of a double-barrel ...

journal_title：Nature protocols

authors： Sylantyev S,Rusakov DA

更新日期：2013-01-01 00:00:00

abstract：:The tracking of cell fate, shape and migration is an essential component in the study of the development of multicellular organisms. Here we report a protocol that uses the protein Kaede, which is fluorescent green after synthesis but can be photoconverted red by violet or UV light. We have used Kaede along with confo...

journal_title：Nature protocols

authors： Hatta K,Tsujii H,Omura T

更新日期：2006-01-01 00:00:00

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

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

更新日期：2015-12-01 00:00:00

abstract：:Boronic acids and esters have critical roles in the areas of synthetic organic chemistry, molecular sensors, materials science, drug discovery, and catalysis. Many of the current applications of boronic acids and esters require materials with very low levels of transition metal contamination. Most of the current metho...

journal_title：Nature protocols

authors： Mfuh AM,Schneider BD,Cruces W,Larionov OV

更新日期：2017-03-01 00:00:00

abstract：:As proteins within cells are spatially organized according to their role, knowledge about protein localization gives insight into protein function. Here, we describe the LOPIT technique (localization of organelle proteins by isotope tagging) developed for the simultaneous and confident determination of the steady-stat...

journal_title：Nature protocols

authors： Sadowski PG,Dunkley TP,Shadforth IP,Dupree P,Bessant C,Griffin JL,Lilley KS

更新日期：2006-01-01 00:00:00

abstract：:A major challenge in the analysis of gene expression microarray data is to extract meaningful biological knowledge out of the huge volume of raw data. Expander (EXPression ANalyzer and DisplayER) is an integrated software platform for the analysis of gene expression data, which is freely available for academic use. It...

journal_title：Nature protocols

authors： Ulitsky I,Maron-Katz A,Shavit S,Sagir D,Linhart C,Elkon R,Tanay A,Sharan R,Shiloh Y,Shamir R

更新日期：2010-02-01 00:00:00