isolation Protocols

Category: RNA Protocols: RNA-binding Protein Purification

A high yield affinity purification method for specific RNA-binding proteins: isolation of the iron regulatory factor from human placenta. Neupert 1990. - [Read A high yield affinity purification method for specific RNA-binding proteins.]

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

The protocol includes: organelle isolation, deoxyribonuclease treatment, lysis, deproteinisation and a final DNA purification with sodium dodecyl sulphate and potassium acetate. The organelle DNA yield is 5–10 micrograms per gram of tissue and the DNA is fully restrictable. The technique is inexpensive and appropriate for the isolation of multiple samples of organelle DNA from a small amount of tissue. - [Read A Method for Isolation of Chloroplast DNA and Mitochondrial DNA from Sunflower]

Category: Molecular Biology Protocols: Carbohydrate Protocols

Solvent partition protocol allows the isolation of gangliosides from small samples and from samples where ganglioside concentrations are low, especially relative to the concentration of potentially contaminating proteins and other large molecular weight species. Stephan Ladisch Director, Center for Cancer and Transplant Biology, Children's National Medical Center, Washington, D.C. - [Read A Method for Micro-Scale Isolation and Purification of Gangliosides]

Category: Fungi Protocols: Aspergillus Protocols

Protocol for the rapid method for isolation of total nucleic acids from Aspergillus nidulans. - [Read A Rapid Method for Isolation of Total Nucleic Acids from Aspergillus nidulans Protocol]

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

Procedure to isolate DNA from Arabidopsis Thaliana. - [Read Arabidopsis Thaliana DNA Isolation Procedure]

Category: Plant Biology Protocols: Arabidopsis Nucleic Acid Protocols

Protocol for Arabidopsis thaliana DNA isolation procedure. - [Read Arabidopsis thaliana DNA Isolation Protocol]

Category: Bacterial Cell Culture Protocols: Bacterial Stocks Protocols

4 strains of E. coli are used in these studies: JM101 for M13 infection and isolation, XL1BMRF'for M13 or pUC-based DNA transformation, and ED8767 for cosmid DNA transformation. To maintain their respective F' episomes necessary for M13 viral infection, JM101 is streaked onto a M9 minimal media plate and XL1BMRF' is streaked onto an LB plate containing tetracycline. ED8767 is streaked onto an LB plate. These plates are incubated at 37degC overnight. For each strain, 3 ml. of appropriate liquid.. - [Read Bacterial Cell Maintenance Protocol]

Category: Cell Biology and Cell Culture

OptiPrep Application sheet C14 describes procedures for determining the density and sedimenting properties of any cell (of any size or density) using either a continuous or discontinuous gradient of iodixanol. This Application Sheet describes procedures aimed at isolating specifically a relatively low-density cell fraction from any tissue. - [Read C25 Isolation from spleen, thymus, pancreas, alveolar tissue and other tissues]

Category: Cell Biology and Cell Culture

The protocol presented in this Application Sheet uses an alternative strategy to sedimentation on to a density barrier, that is to adjust the density of whole blood to a value just greater than the cells of interest and allow them to float to the surface. - [Read C8 Isolation of bovine peripheral blood mononuclear cells by flotation.]

Category: Cell Organelle Protocols: Nucleus Protocols

Cajal Body Isolation Protocol. Protocol includes: Sonication, Removal Nucleoi, Gradient One, Gradient two, Concentration and final enrichment of cajal bodies. Also includes: Making 2.55M sucrose stock and Analysis of the enriched Cajal body fraction. - [Read Cajal Body Isolation Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Sorting Protocols

The combination of prospective identification/isolation of bone marrow progenitors and quantitative RT-PCR is a powerful tool to understand the molecular mechanism underlying hematopoiesis. Describes the standard procedures of the murine myeloid progenitor staining for fluorescence activated cells sorting (FACS) and RNA purification methods. - [Read Cell Staining for Sorting of Hematopoietic Stem Cells (HSC) and Myeloid Progenitors]

Category: Stem Cell Protocols

The combination of prospective identification/isolation of bone marrow progenitors and quantitative RT-PCR is a powerful tool to understand the molecular mechanism underlying hematopoiesis. Here, we described our standard procedures of the murine myeloid progenitor staining for fluorescence activated cells sorting (FACS) and RNA purification methods. - [Read Cell Staining for Sorting of Hematopoietic Stem Cells and Myeloid Progenitors and Isolating RNA]

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

Protocol for chloroplast DNA isolation. - [Read Chloroplast DNA Isolation Protocol]

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

Protocol for the isolation of DNA from the chloroplast. - [Read Chloroplast Isolation DNA Protocol]

Category: Cell Organelle Protocols: Nucleus Protocols

Protocol for chromatin isolation by small-scale biochemical fractionation. - [Read Chromatin Isolation by Small-Scale Biochemical Fractionation Protocol]

Category: DNA Protocols: Cosmid Library Protocols

Protocol describes how to construct a library of 35-45-kb fragments of genomic DNA in the double cos site cosmid vector, SuperCos-1. The steps include: Linearization and dephosphorylation of SuperCos-1 DNA; Partial digestion of high-molecular-weight DNA with MboI; Dephosphorylation of high-molecular-weight genomic DNA; Ligation of cosmid arms to genomic DNA: Packaging and plating recombinants; Isolation and analysis of recombinant cosmids: Validation of the library. - [Read Construction of Genomic DNA Libraries in Cosmid Vectors Protocol]

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

The starting material for de novo isolation of stem cell lines can be either normal 3.5-days post coitum (dpc) expanded blastocysts or "delayed" blastocysts. Delayed blastocysts are usually collected 4-6 days after ovariectomy. For both groups of blastocysts, tissue culture procedures are similar. The only difference is the timing of the first disaggregation, because delayed blastocysts will initially grow more slowly. - [Read De Novo Isolation of Embryonic Stem (ES) Cell Lines from Blastocysts Protocol]

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

Protocol for isolation of dendritic cell derived exosomes. - [Read Dendritic Cell Derived-Exosomes]

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

A protocol for extraction or isolation of both DNA and RNA from the same material, typically plant leaf or leaves. - [Read DNA and RNA Extraction Protocols]

Category: DNA Protocols: DNA Extraction Blood

Purifying nuclear pellets, Extraction of DNA from nuclei. Method for DNA isolation from Blood. Genomic Variation Laboratory, UC Davis. - [Read DNA extraction from blood]

Category: DNA Protocols: DNA Extraction Blood

Method for isolation of DNA From Whole Blood for PCR. The Jackson Laboratory. - [Read DNA From Whole Blood for PCR]

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: Genomic DNA Isolation Protocols

In this protocol, the DNA-binding capacity of Wizard MagneSil particles is used to capture and release a consistent amount of DNA (100 ng) across a wide range of samples.  At the end of the procedure, the DNA is eluted into 100 µl Elution Buffer to give a final concentration of 1 ng/µl, relieving the need for postpurification DNA quantitation. - [Read DNA IQ Isolation of Genomic DNA from Stains and Buccal Swabs Protocol]

Category: BACs Bacterial Artificial Chromosome Protocols

Rapid alkaline lysis miniprep method for isolating DNA from large PAC clones. It is a modification of a standard Qiagen-Tip method that uses no organic extractions or columns. The method works very well for doing analytical restriction digests of PAC clones and can be scaled up if necessary. - [Read DNA Isolation From BAC & PAC Clones Protocol]

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols

This is a rapid alkaline lysis miniprep method for isolating DNA from large PAC clones.  It is a modification of a standard Qiagen-Tip method that uses no organic extractions or columns.  The method works very well for doing analytical restriction digests of PAC clones and can be scaled up if necessary. - [Read DNA Isolation From BAC & PAC Clones Protocol]

Category: DNA Protocols: DNA Extraction Protocols

Allows students to easily isolate chromosomal DNA from onion cells using the same basic tools and methods that scientists use in the lab. A good introduction to using pipettes. Kate Dollard - [Read DNA Isolation from Onion Cells]