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CaCl2 Competent Cell Stock Protocol - http://www.bio.indiana.edu/~chenlab/potocols/CompetentCellStock.htm

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Simple Protocol for CaCl2 Competent Cells. Chen Lab. Indiana University Bloomington - [Read CaCl2 Competent Cell Stock Protocol]

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 Cell Preparation [CaCl2] - http://www.cf.ac.uk/biosi/staff/kille/Methods/Gene/Comp_cell_CaCl.html

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Nice protocol for the preparation of Bacterial cells to allow uptake of plasmid vectors. Dr Peter Kille. Hot Metals Cardiff. - [Read Competent Cell Preparation [CaCl2]]

Procedure for Transfection of Mammalian Cells Using Lipofectamine - http://www.research.umbc.edu/~jwolf/m10.htm

Category: Cell Biology and Cell Culture: Cell Transfection Protocol

Lipofectamine transient tranfection of mammalian cells in IMDM cell media. Julie B. Wolf, UMBC. - [Read Procedure for Transfection of Mammalian Cells Using Lipofectamine]

Fixation Protocol - http://cmif.osu.edu/419.cfm

Category: Histology Protocols: Fixation of Cells Tissues Protocol

There are many variations based on the type of tissue to be examined, whether one is dealing with cell suspensions, biopsies, perfused tissues, or monolayer . Fixation Protocol. OSU Campus Microscopy & Imaging Facility :: The Ohio State University College of Med - [Read Fixation Protocol]

Cell Staining for Immunofluorescence Microscopy - http://www.bdbiosciences.com/pharmingen/protocols/Immunofluorescence_Microscopy.shtml

Category: Molecular Imaging: Immunofluorescence Microscopy

Cell Staining for Immunofluorescence Microscopy. Includes protocols for fixing the cells, Coverslip Preparation for Adherent Cells, Coverslip Preparation for Non-Adherent Cells, Paraformaldehyde Fixation, and Methanol/Acetone Fixation. Blocking protocols include blocking with primary antibody, and incubation with secondary antibody. - [Read Cell Staining for Immunofluorescence Microscopy]

Handbook of Cell Culture. - Sigma Aldrich. - http://www.sigmaaldrich.com/Area_of_Interest/Life_Science/Cell_Culture/Key_Resources/ECACC_Handbook/Cell_Culture_Techniques_12.html

Category: Cell Biology and Cell Culture: Cell Culture Techniques

ECACC Handbook of Cell Culture. Fundamental Techniques in Cell Culture: A Labratory Handbook. Sigma Aldrich. Excellent Guide to Cell Culture, aseptic technique, storage of cell lines, subculture of cells. - [Read Handbook of Cell Culture. - Sigma Aldrich.]

Cell Lysate Extracts - http://www.upstate.com/misc/protocol_detail.q.prot.e.CellLysate_Prep.a.name.e.Cell_Lysate_Extracts-General_Protocols

Category: Protein Protocols: Protein Extraction From Cells

Cell Lysate Extracts. Great protocols for cell lysis preparation from a variety of cell types. There are numerous methods of cell stimulation and lysis. For a given protein, Upstateâ€™s Laboratories determine the specific treatment upon initial testing of its products. It is important to select the correct cell line, stimulation procedure (if any), and lysis protocol. Upstate. - [Read Cell Lysate Extracts]

Cell Fixation Using Methanol - http://www.lamondlab.com/pdf/CellFixMEOH.pdf

Category: Immunology: Immunocytochemistry Protocols

Methanol Fixation for Immunofluorescence. Lamond Lab, Univ. Dundee - [Read Cell Fixation Using Methanol]

Western Blot Detailed (PDF) - http://www.its.caltech.edu/%7Ebjorker/Western_blot.pdf

Category: Protein Protocols: Western Blot Protocol

Protocol includes cell lysate preparation, SDS-PAGE gels, transfer, and blotting. (Bjorkman Group). - [Read Western Blot Detailed (PDF)]

Hippocampal Neuron Cell Cultures - http://www.pharm.stonybrook.edu/faculty/tsirka/lab/HippocampalNeuronCultures.doc

Category: Neuroscience: Neuron Cell Culture Protocols

Hippocampal Neuron Culture Protocol, methods. Jean Siao, Ph.D. Tsirka Lab. - [Read Hippocampal Neuron Cell Cultures]

Large scale nuclear extract preparation - http://mesentsev.narod.ru/Biblioteque/Hattori.html

Category: Protein Protocols: Nuclear Extraction Protocols

Homogenize cells or tissue, Pellet nuclei, Lyse nuclei, Reprecipitate nuclear proteins, dialyze nuclear extract. Hattori M et al., DNA Cell Biol. 1990. Homogenization buffer and dialysis buffer recipes. - [Read Large scale nuclear extract preparation]

Dose of G418 to be used for selection - http://invitrogen.custhelp.com/cgi-bin/invitrogen.cfg/php/enduser/std_adp.php?p_sid=rLTNWTng&p_lva=&p_faqid=80

Category: Cell Biology and Cell Culture: Cell Transfection Protocol

What dose of Geneticin (G418) should I use for stable cell line selection? Invitrogen FAQs. - [Read Dose of G418 to be used for selection]

Hanahan Competent Cell Protocol. - http://www.rockefeller.edu/labheads/vosshall/protocols/Hanahan.pdf

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Hanahan Competent Cell Protocol. PROTOCOL PRODUCES ABOUT 160 X 250ul ALIQUOTS OF COMPETENT CELLS. Vosshall. Rockefeller. - [Read Hanahan Competent Cell Protocol.]

Preparation of Competent Bacterial Cells - http://www.cbs.umn.edu/~amundsen/chlamy/methods/competent.html

Category: Microbiology: Bacterial Competent Cells Preparation E.Coli

Simple E.coli competent cell Protocol. Based on a protocol from Neil Olszewski and Scott Medberry.. Craig D. Amundsen - [Read Preparation of Competent Bacterial Cells]

PFA Fixation for Immunofluorescence - http://www.lamondlab.com/pdf/Cellfixpfa.pdf

Category: Immunology: Immunocytochemistry Protocols

Cell fixation using paraformaldehyde. - [Read PFA Fixation for Immunofluorescence]

HISTOLOGY-RELATED PROTOCOLS Fixation and Frozen Block PDF - http://www.uchsc.edu/misc/diabetes/derc/protocols/histologyprotocols.pdf

Category: Histology Protocols: Fixation of Cells Tissues Protocol

Protocols for cell fixation and tissue fixation. HISTOLOGY-RELATED PROTOCOLS Fixation and Frozen Block techniques. Embedding Tissue Into Frozen Blocks. Fixing Cells with Fresh Paraformaldehyde. Preparation of Frozen Sections. Immunohistochemistry. Paraffin sections and Frozen sections. DERC Univ. Colorado PDF. - [Read HISTOLOGY-RELATED PROTOCOLS Fixation and Frozen Block PDF]

Extraction and Purification of Total RNA using TRIzol OR TRI Reagent Protocol - http://www.animal.ufl.edu/hansen/Protocols/RNA_extraction.htm

Category: Nucleic Acid Purification Protocols: RNA Isolation Protocols

Protocol for extraction and purification of total RNA using TRIzol OR TRI reagent. Includes: Homogenization for Cell Suspensions; Phase Separation; RNA Precipitation; RNA Wash; Redissolving the RNA; Determination of RNA Concentration and Purity; Preparation of Rnase-free water. - [Read Extraction and Purification of Total RNA using TRIzol OR TRI Reagent Protocol]

TCA/Acetone Precipitation (Large Scale) PDF - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000148.pdf?pid=PP00000148

Category: Protein Protocols: Protein Precipitation Procols

A protein precipitation procedure used to precipitate protein from cell lysates. Allows for optimal protein recovery and accurate assays. The Signaling Gateway - [Read TCA/Acetone Precipitation (Large Scale) PDF]

Basic Techniques for Mammalian Cell Tissue Culture Protocol - https://catalog.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeID=9E662B5FE053DEE8CB4AC3EB6F729A11&objectid=667396C6F0D96C40EB68A3D8DB8B201A

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

Cultured mammalian cells are used extensively in cell biology studies; it requires a number of special skills in order to be able to preserve the structure, function, behavior and biology of the cells. This unit describes the basic skills required to maintain and preserve cell cultures: aseptic technique, medium characteristics, passaging, freezing and storage, recovering frozen stocks, and counting viable cells. - [Read Basic Techniques for Mammalian Cell Tissue Culture Protocol]

Calcium Phosphate Transfection Method - http://www.flemingtonlab.com/Protocols/CalciumPhosphateTransf.pdf

Category: Transfection Protocols: Calcium Phosphate Transfection Protocols

This calcium phosphate transfection method works best in cell lines that are 1) highly transformed and 2) adherent (Hela, U2OS, SAOS2, AdAH, NPC-KT and obtain from 20% to 100% transfection efficiency depending on the cell line). Works well for transient experiments but precautions should be used in the design and interpretation of experiments based on the discussion below. Also works very well for generating stable cell lines. This method is quite sensitive to the amount of input plasmid. - [Read Calcium Phosphate Transfection Method]

Cell Thawing and cell line Freezing Protocol - http://iacf.bsd.uchicago.edu/Monoclonal_Antibody_Facility/FreezeThawProtocol.htm

Category: Cell Biology and Cell Culture: Cell Culture Techniques: Cell Line Preservation Freezing

Method for freezing and thawing cells. Immunology Applications Core Facility. U. Chicago - [Read Cell Thawing and cell line Freezing Protocol]

Introduction to Animal Cell Culture - Corning Life Science PDF. - http://www.protocol-online.org/cgi-bin/prot/jump.cgi?ID=3309

Category: Cell Biology and Cell Culture: Cell Culture Techniques

Introduction to Animal Cell Culture. Corning Life Science PDF. Key concepts to Cell Cultures an Introductory Manual with definitions and explanations. - [Read Introduction to Animal Cell Culture - Corning Life Science PDF.]

Electroporation of Cell Lines With DNA - http://users.path.ox.ac.uk/~ciu/Methods/DNA/electroporation.html

Category: Cell Biology and Cell Culture: Cell Electroporation Protocols

Electroporation of Cell Lines With DNA. Electroporation for the efficient transfection of mammalian cells with DNA. Chu et al. Cellular Immunology Oxford - [Read Electroporation of Cell Lines With DNA]

MTT Cell Proliferation Assay Protocol - http://www.atcc.org/pdf/30-1010k.pdf

Category: Cell Biology and Cell Culture: Cell Viability Protocols

The MTT Cell Proliferation Assay measures the cell proliferation rate and conversely, when metabolic events lead to apoptosis or necrosis, the reduction in cell viability. The number of assay steps has been minimized as much as possible to expedite sample processing. The MTT Reagent yields low background absorbance values in the absence of cells. Includes: Determining optimal cell counts, performing an assay, data interpretations and troubleshooting. - [Read MTT Cell Proliferation Assay Protocol]

Articles (1-10 of 28)

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.

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.

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.

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.

Microinjection Pipettes Protocol

Microinjection Pipettes Preparation Protocol.

Acid Washing Microinjection Pipettes Protocol

Acid Washing of Glass microinjection pipettes protocol.

Paraffin Embedding Protocol

Paraffin Embedding Protocol for molecular profiling. This Paraffin Embedding Protocol describes the processing of the tissues into sections following ethanol fixation. Molecular profiling (MP) is a technique that is used to visualize the global patterns of RNA expression or protein expression in various cell types and disease processes.

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