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CALCIUM PHOSPATE TRANSFECTION OF HUMAN CELLS - http://omrf.ouhsc.edu/~frank/CAPO4TRANSFECT.html

Category: Cell Biology and Cell Culture: Cell Transfection Protocol

Simple Transfection using Calcium chloride and phosphate.Dr. Frank. Arthritis and Immunology Program, Oklahoma City. Medical Research Foundation. - [Read CALCIUM PHOSPATE TRANSFECTION OF HUMAN CELLS]

Competent cell preparation - http://www.chem.uga.edu/scottgrp/GrpProtocols/Competent_cell_preparation.htm

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Bacterial E.Coli competent cell preparation. Calcium Chloride Protocol, Preparation of calcium chloride competent cells for frozen storage, Electroporation Protocol, Electroporation Protocol for transformations using double-stranded plasmids, Reference: - [Read Competent cell preparation]

Competent cells, calcium chloride method, E. coli, description - http://web.uct.ac.za/depts/mmi/bbhelp/bac1.html

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Competent cells, calcium chloride method, E. coli, description. CaCl2 and DMSO methods for preparing bacterial and E.Coli competent cells. Zappe, H. BBHelp. - [Read Competent cells, calcium chloride method, E. coli, description]

Fixation. - http://www.dakousa.com/ishbfixation.pdf

Category: Histology Protocols: Histological Fixation

FORMALDEHYDE-BASED FIXATIVES, PARAFFIN-EMBEDDED SECTIONS, BOUINâ€™S SOLUTION, MERCURIC CHLORIDE-BASED FIXATIVES. FARMILO et al. - [Read Fixation.]

How to Make Competent Cells - Calcium Chloride - http://sosnick.uchicago.edu/competent_cells.html

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

How to Make Competent Cells - Calcium Chloride. Sosnick Lab, Univ. Chicago - [Read How to Make Competent Cells - Calcium Chloride]

Hybridization of Oligonucleotide Probes in Aqueous Solutions Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3883

Category: Nucleic Acid Modification Protocols: Nucleic Acid Labeling Protocols

Hybridization is carried out in conventional aqueous solvents at a temperature well below the predicted melting temperature. Nonspecific hybrids are then removed by washing at high stringency in buffers containing quaternary salts. Tetramethylammonium chloride (TMACl) is used with probes that are 14-50 nucleotides in length, whereas tetraethylammonium chloride (TEACl) is used with longer oligonucleotides. - [Read Hybridization of Oligonucleotide Probes in Aqueous Solutions Protocol]

Preparation and Transformation of Competent E. coli Using Calcium Chloride Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3932

Category: Bacteria Transformation Protocols

Protocol used to prepare batches of competent bacteria that yield 5 x 106 to 2 x 107 transformed colonies/µg of supercoiled plasmid DNA. - [Read Preparation and Transformation of Competent E. coli Using Calcium Chloride Protocol]

Preparation and Transformation of Competent E. coli Using Calcium Chloride Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3932

Category: E Coli Protocols: E coli Transformation Protocols

Protocol is is used to prepare batches of competent bacteria that yield 5 x 106 to 2 x 107 transformed colonies/µg of supercoiled plasmid DNA. - [Read Preparation and Transformation of Competent E. coli Using Calcium Chloride Protocol]

Preparation of RNA from Plant Tissue Using Guanidinium Isothiocyanate/Cesium Chloride Ultracentrifug - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4102

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

Tissues are homogenized in guanidinium isothiocyanate lysis buffer and then subjected to cesium chloride ultracentrifugation. The resulting RNA is then extracted with phenol and chloroform to remove protein and lipid contaminants. - [Read Preparation of RNA from Plant Tissue Using Guanidinium Isothiocyanate/Cesium Chloride Ultracentrifug]

Purification of Plasmid DNA by Precipitation with Polyethylene Glycol Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3907

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: DNA Plasmid Preps Protocols

Crude preparations of plasmid DNA are first treated with lithium chloride and Rnase (to remove RNA). The plasmid DNA is then precipitated in a solution containing polyethylene glycol and MgCl2. - [Read Purification of Plasmid DNA by Precipitation with Polyethylene Glycol Protocol]

Real-Time RT-PCR: cDNA Synthesis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4114

Category: PCR Protocols: Real-Time PCR Protocols

Protocol uses the Superscript II First-Strand Synthesis system for the generation of cDNA from total RNA. RNA purified using TRIzol reagent (Invitrogen) or the methods described in Preparation of RNA Using Guanidinium Isothiocyanate/Cesium Chloride Ultracentrifugation, Preparation of RNA from Paraffin-Embedded Fixed Tissue. - [Read Real-Time RT-PCR: cDNA Synthesis Protocol]

Real-Time RT-PCR: cDNA Synthesis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4114

Category: PCR Protocols: Reverse Transcriptase RT-PCR Protocols

Removal of Small Fragments of Nucleic Acid from Preparations of Plasmid DNA by Centrifugation - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3916

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: DNA Plasmid Preps Protocols

Contamination of plasmid DNA by fragments of DNA and RNA is reduced to an acceptable level by centrifugation through 1 M sodium chloride. This method was devised by Brian Seed when he was a graduate student at Harvard University. - [Read Removal of Small Fragments of Nucleic Acid from Preparations of Plasmid DNA by Centrifugation]

Staining of Nerve Fibers Protocol - http://library.med.utah.edu/WebPath/HISTHTML/MANUALS/BODIAN.PDF

Category: Neuroscience Protocols: Neuronal Stains Protocols

Protargol-S (silver proteinate) is used with the addition of copper metal. The copper replaces the silver in the connective tissue, allowing a greater differentiation between the nerve fibers and the connective tissue. The silver is reduced with hydroquinone to the visible metallic form. The sections are toned with gold chloride, the gold chloride is reduced with oxalic acid, increasing the deposit of metallic gold on the sections. - [Read Staining of Nerve Fibers Protocol]

Transformation of Aspergillus niger Protocol - http://clone.concordia.ca/aspergillus/protocols/nigtransf.html

Category: Fungi Protocols: Aspergillus Protocols

Protocol for the transformation of Aspergillus niger. This procedure is done by first digesting the outer cell wall, forming protoplasts, and then by making holes in the membrane through which the dna can enter using calcium chloride and polyethylene glycol. Includes: Protocol for making A.niger protoplasts; Transformation; Plating. - [Read Transformation of Aspergillus niger Protocol]

Articles (1-10 of 15)

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.

Drosophila Cell Transfection Protocol

A simple Drosophila tissue culture cell transfection method.

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.

Probe Preparation for 22 x 40 mm DNA Microarrays

Probe Preparation for 22 x 40 mm DNA Microarrays Protocol.

In Vitro Translated Xenopus Mos Kinase Assay Protocol

In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.

Antibodies Phage Expression Protocol Using 96-well Microtiter Plates

A Phage display expression protocol for antibody expression in 96-well microtiter plates.

Immobilized Antigen Phage Antibody Selection Protocol

A protocol for the selection of Phage Antibodies using Immobilized Antigen. This method describes the selection of antibodies from bacteriophage antibody libraries that recognize a specific antigen. The phage display library of antibody-displaying phage particles is exposed to antigen attached to a solid substrate (Nunc Immuno™ tubes). The phage particles with affinity for antigen bind to the immobilized antigen and are selected from the library of phage expressing antibodies.

Coupling Cysteine Containing Peptides to Carrier Protein for Immunization and Polyclonal Antibody

This protocol is used in the production of polyclonal antibodies that recognize a peptide sequence of interest.

Absorption Spectroscopy and Quantification of Filamentous Phage Protocol

Unlike spherical phage, such as T4 and λ, which have roughly equal weight ratios of protein to DNA, filamentous phage have about six times more protein than DNA; the protein therefore contributes substantially to the absorption spectrum.

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