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1,115 Newest Publications in csh protocols

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Rapid Isolation of Yeast DNA

01-Jun-2018 | Green, M. R., Sambrook, J., CSH Protocols, 2018

This method is used to isolate yeast genomic DNA or shuttle plasmids that replicated both in Saccharomyces cerevisiae and in E. coli.

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Analysis of Cell Viability by the MTT Assay

01-Jun-2018 | Kumar, P., Nagarajan, A., Uchil, P. D., CSH Protocols, 2018

Among viability assays that depend on the conversion of substrate to chromogenic product by live cells, the MTT assay is still among one of the most versatile and popular assays. The MTT assay involves the conversion of the water-soluble yellow dye MTT ...

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Assembly of Spindles and Asters in Xenopus Egg Extracts

01-Jun-2018 | Field, C. M., Mitchison, T. J., CSH Protocols, 2018

Here, we provide methods for assembly of mitotic spindles and interphase asters in Xenopus laevis egg extract, and compare them to spindles and asters in the egg and zygote. Classic "cycled" spindles are made by adding sperm nuclei to metaphase-arrested cytostatic factor (CSF) extract and ...

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Preparation of Antibody Using Caprylic Acid

01-Jun-2018 | Fishman, J. B., Berg, E. A., CSH Protocols, 2018

Caprylic acid has been used to enrich IgG from serum, ascites, and cell culture supernatants by precipitating the non-IgG serum proteins. By precipitating all of the unwanted serum proteins rather than the antibodies, the tendency of antibodies to aggregate when precipitated is avoided. This ...

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ChIP-seq

01-May-2018 | Kim, T. H., Dekker, J., CSH Protocols, 2018

Owing to its digital nature, ChIP-seq has become the standard method for genome-wide ChIP analysis. Using next-generation sequencing platforms (notably the Illumina Genome Analyzer), millions of short sequence reads can be obtained. The densities of recovered ChIP sequence reads along the genome ...

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The Basic Polymerase Chain Reaction (PCR)

01-May-2018 | Michael R. Green; Joseph Sambrook, CSH Protocols, 2018

This protocol describes the reagents and procedures required to amplify a segment of double-stranded DNA in a chain reaction catalyzed by a thermostable DNA polymerase. It is the foundation for all subsequent variations of the polymerase chain reaction (PCR).

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Isolation of DNA from Mouse Tails without Extraction by Organic Solvents

01-May-2018 | Green, M. R., Sambrook, J., CSH Protocols, 2018

This protocol describes a variation of a simple method for isolation of DNA from mouse tails that does not require extraction with phenol:chloroform. This variant is useful when processing very large numbers of samples.

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Fluorescence In Situ Hybridization of Cryosectioned Xenopus Oocytes

01-May-2018 | Neil, C. R., Mowry, K., CSH Protocols, 2018

Xenopus laevis oocytes are widely used to study mechanisms of RNA function and biogenesis. While the large size of Xenopus oocytes is amenable to both biochemical and imaging approaches, the relative opacity of the yolk-rich cytoplasm has limited high-resolution imaging of endogenous RNAs. Here, ...

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Making Weak Antigens Strong: Modifying Antigens by Dinitrophenol or Arsynyl Coupling

01-May-2018 | Greenfield, E. A., DeCaprio, J., Brahmandam, M., CSH Protocols, 2018

Many compounds on their own do not have all of the properties needed to induce a strong antibody response. However, small changes in the structure of an antigen can often greatly alter the immunogenicity of a compound. Common methods for doing so include the addition of small modifying groups ...

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Mouse Embryo Cryopreservation by High-Osmolality Vitrification

01-May-2018 | Mochida, K., CSH Protocols, 2018

In conventional vitrification methods, the embryos are vitrified under considerable supercooling (i.e., under nonequilibrium conditions). This protocol is a refinement of newer equilibrium vitrification methods. The equilibrium vitrification solution contains a high concentration of ...

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