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Bacterial Glycerol Stocks Protocol - http://www.unc.edu/depts/van_dyke/protocols/bacteria/bacterial_glycerol_stocks.pdf

Category: Bacterial Cell Culture Protocols: Bacterial Stocks Protocols

To 2mls of mid-log culture or 1ml of freshly saturated culture add 1ml(or an equal volume) of glycerol solution, mix gently, then freeze rapidly in liquid nitrogen or on dry ice. Store at -20°C and - 70°C. - [Read Bacterial Glycerol Stocks Protocol]

Estimation of Cell Number by Hemocytometry Counting Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4454

Category: Cell Culture Protocols

Protocol describes a method for estimation of mammalian cell number in a defined volume of medium using a hemocytometer. Automated methods using cell-counting devices such as those produced by Coulter are desirable when large numbers of individual samples are to be counted. - [Read Estimation of Cell Number by Hemocytometry Counting Protocol]

Feeder Cell Density - A key Parameter in Human Embryonic Stem Cell Culture - http://www.bioone.org/bioone/?request=get-document&issn=1543-706X&volume=040&issue=08&page=0255

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: ES Cell Growth Media and Requirements

Feeder Cell Density - A key Parameter in Human Embryonic Stem Cell Culture - [Read Feeder Cell Density - A key Parameter in Human Embryonic Stem Cell Culture]

Hydrodynamics-Based Gene Transduction Protocol - http://www.natureprotocols.com/2006/06/29/hydrodynamicsbased_gene_transd.php

Category: Gene Delivery Protocols

This highly efficient in vivo gene transduction technique for laboratory mice. Hepatocytes are most effectively transduced by tail vein injection of a large volume of DNA solution in a short time. Practice with the injection technique is necessary!!! - [Read Hydrodynamics-Based Gene Transduction Protocol]

Immunoblotting: Submerged Electrophoretic Transfer of Proteins from Gels to Membranes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/16/pdb.prot4302

Category: Western Blot Protocols: General Western Blot Methods Protocols

Transfer of proteins from a Tris/glycine SDS-polyacrylamide gel to a membrane using the submerged method is achieved by placing the gel next to a piece of nitrocellulose filter, submerging this sandwich in a large volume of transfer buffer in a transfer tank, and running current from one side of the transfer tank to another. The proteins are then eluted by transferring them from the gel onto the filter. - [Read Immunoblotting: Submerged Electrophoretic Transfer of Proteins from Gels to Membranes Protocol]

Isolation of DNA from Mammalian Cells by Spooling Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4037

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: DNA Isolation from Mammalian Cells Protocols

Procedure is used to prepare DNA simultaneously from many different types of samples or tissues. Although the DNA is generally too small (approx. 80 kb) for efficient construction of genomic DNA libraries, it gives excellent results in Southern hybridizations and PCRs. Cultured aneuploid mammalian cells (2 x 107, e.g., HeLa cells) yield 100 µg of DNA in a volume of 1 ml. - [Read Isolation of DNA from Mammalian Cells by Spooling Protocol]

List of Protein Assays - http://www.ruf.rice.edu/~bioslabs/methods/protein/protein.html

Category: Protein Protocols: Protein Quantitation Concentration Protocols

Find a list of assays for the determination of protein concentration in a solution. This list includes the sensitivity range, volume/amount of sample needed, subjective comments on accuracy and convenience, and major interfering agents. Procedural details, equipment requirements, and references are outlined in the individual assay documents. - [Read List of Protein Assays]

Measuring Volume Protocol - http://homepages.gac.edu/~cellab/chpts/chpt1/ex1-6.html

Category: Microscopy Protocols

Protocol for measuring volume. Material required includes: Microscope, Hemacytometer and coverslip, Suspension of yeast. - [Read Measuring Volume Protocol]

Microinjection of Protein Samples Protocol - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4657

Category: Microinjection Protocols: Microinjection of Proteins Protocols

This protocol describes a method for microinjecting proteins into the nucleus or cytoplasm of adherent cells. Microinjection equipment can be obtained from a number of suppliers; this protocol has been used with the Narishige IM-200 air pressure regulator and the Leitz micromanipulator. Using this system, it is possible to microinject a constant volume within a 50% difference among cells. - [Read Microinjection of Protein Samples Protocol]

Optimizing Electrotransfection of Mammalian Cells In Vitro Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/1/pdb.prot4449

Category: Transfection Protocols

Protocol describes transfection of plasmid DNA into mammalian cell lines using electroporation, a process whereby external application of electric pulses induce cell membrane permeability. Cells in suspension and small volume cells are difficult to transfect, whereas adherent cells and large volume cells are relatively easy. Regardless of cell size or phenotype, transfection efficiency increases with a high concentration of cells in a small volume. - [Read Optimizing Electrotransfection of Mammalian Cells In Vitro Protocol]

Optimizing Electrotransfection of Mammalian Cells In Vitro Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/1/pdb.prot4449

Category: Transfection Protocols: Electroporation Sonoporation Protocols

Protocol describes transfection of plasmid DNA into mammaliancell lines using electroporation, a process whereby externalapplication of electric pulses induce cell membrane permeability.A number of factors can affect electrotransfection efficiency.In general, cells in suspension and small volume cells are difficultto transfect, whereas adherent cells and large volume cellsare relatively easy. - [Read Optimizing Electrotransfection of Mammalian Cells In Vitro Protocol]

Serum Testing for Mammalian Cell Culture Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4348

Category: Primary Cell Isolation and Culture Protocols: Mammalian Cell Culture Protocols

This protocol describes a method for testing of new serum lots prior to use in cell culture. Serum lots vary considerably in their ability to support cell growth, and some lots even contain toxic or growth-inhibitory compounds. It is advantageous to test serum lots and purchase in large volume, both for cost benefit. - [Read Serum Testing for Mammalian Cell Culture Protocol]

Single Strand DNA Prep For Sequencing Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/S2.html

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols

Protocol for single strand DNA prep for sequencing. This protocol works well from a 5 ml starting culture or a 1 ml starting culture (adjust volume accordingly). - [Read Single Strand DNA Prep For Sequencing Protocol]

Articles (1-10 of 18)

Populational Analysis of Genomic DNA Protocol

A protocol on the populational Analysis of genomic DNA.

Preparing Stock Plating of Bacteria Protocol

A protocol for the preparation of stock plating of bacteria.

Lambda Phage Protocol for Large Preparation DNA Recovery

Lambda Phage Protocol for Large Preparations DNA Recovery.

Lambda Phage Protocol Large DNA Preparation

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

Genomic DNA Labeling of Microarrays Protocol

DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.

Tubulin Polymerization Protocol using GTP and GMPCPP

Tubulin is polymerized into microtubules by incubating tubulin at 37°C with GTP. A nucleation seed is added when the purpose is to assay microtubule elongation. Tubulin can also be polymerized for the purposes of recycling the tubulin or labeling the microtubules with fluorescently labeled tubulin. Based on the protocol by Timothy Mitchison of Harvard University.

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.

Probe Preparation for 22 x 40 mm DNA Microarrays

Probe Preparation for 22 x 40 mm DNA Microarrays Protocol.

Acid Guanidinium Thiocyanate RNA Isolation Phenol Chloroform Extraction

A single step RNA isolation protocol using Phenol Chloroform Extraction and Acid Guanidinium Thiocyanate. This RNA isolation method uses the fact that guanidinium thiocyanate can simultaneously lyse the cells and inactive cellular RNAses during the initial RNA isolation step allow a single step in the method.

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