eukaryotic Protocols

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Electroporation of Eukaryotic Cells - http://omrf.ouhsc.edu/~frank/EPOREUK.html

Category: Cell Biology and Cell Culture: Cell Electroporation Protocols

The protocol uses a Bio-Rad Gene Pulser II apparatus to electroporate DNA into human cell lines. Dr. Frank, Arthritis and Immunology Program, Oklahoma City Medical Research Foundation - [Read Electroporation of Eukaryotic Cells]

Calcium-phosphate-mediated Transfection of Eukaryotic Cells with Plasmid DNAs - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3871

Category: Transfection Protocols: Calcium Phosphate Transfection Protocols

Calcium phosphate forms an insoluble precipitate with DNA, which attaches to the cell surface and is taken into the cells by endocytosis. The protocol is easily adapted for use with other types of cells, both adherent and nonadherent. This protocol is a modified version of a method published by Jordan et al. (1996) who rigorously optimized calcium-phosphate-based transfection methods for Chinese hamster ovary cells and the 293 line of human embryonic kidney cells. - [Read Calcium-phosphate-mediated Transfection of Eukaryotic Cells with Plasmid DNAs]

Freezing and Thawing Eukaryotic Cells - http://omrf.ouhsc.edu/~frank/EukFreeze.html

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

Freezing and Thawing Eukaryotic Cells in liquid nitrogen, Freezing Cell Lysates for Western Blots, Thawing Cells from Liquid Nitrogen. Dr. Bart Frank Oklahoma. - [Read Freezing and Thawing Eukaryotic Cells]

A practical guide to DNA methylation Promega PDF - http://www.promega.com/pnotes/42/methv3/Methv3_core.pdf

Category: Epigenetics and Methylation Protocols

The pattern and extent of DNA methylation can significantly affect the success of restriction digestions or bacterial transformations. Prokaryotic DNA may be methylated by host restriction/modification systems, while eukaryotic DNA often is methylated i - [Read A practical guide to DNA methylation Promega PDF]

Defrosting Eukaryotic Cells Frozen In Liquid N2 Protocol - http://www2.umdnj.edu/~geller/lab/thawing.html

Category: Cell Culture Protocols: Cell Storing and Cell Freezing Protocols

Protocol for the defrosting of eukaryotic cells frozen in liquid N2. - [Read Defrosting Eukaryotic Cells Frozen In Liquid N2 Protocol]

Eukaryotic In Vitro Translation Systems - http://www.promega.com/paguide/chap5.htm#title2

Category: Cell Biology and Cell Culture: In Vitro Reconstitution Protocols

Provides information on specific parameters you need to be aware of when using eukaryotic in vitro translation systems. Includes: DNA Template Considerations; Protein Labeling; Non-Radioactive Protein Labeling. - [Read Eukaryotic In Vitro Translation Systems]

Amino-allyl Labeling - http://pga.tigr.org/sop/M004_1a.pdf

Category: DNA Microarray Protocols

This protocol describes the labeling of eukaryotic RNA with aminoallyl labeled nucleotides via first strand cDNA synthesis followed by a coupling of the aminoallyl groups to either Cyanine 3 or 5 (Cy 3/Cy5) fluorescent molecules. Hasseman. TIGR Microarr - [Read Amino-allyl Labeling]

Construction of cDNA Libraries in Eukaryotic Expression Vectors for Construction and Screening - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4068

Category: DNA Protocols: Genomic DNA Library Protocols

This stage summarizes how to construct a cDNA library in a mammalian vector, using commercial kits. These kits are available from several manufacturers and generally include all the required reagents. - [Read Construction of cDNA Libraries in Eukaryotic Expression Vectors for Construction and Screening]

MICROARRAY PCR, PURIFICATION, AND STORAGE - http://pga.tigr.org/sop/M002_1a.pdf

Category: DNA Microarray Protocols

This protocol describes PCR amplification of eukaryotic cDNA plasmid inserts, gel electrophoresis, purification, and storage of PCR products. Hasseman. TIGR Microarray Protocols. - [Read MICROARRAY PCR, PURIFICATION, AND STORAGE]

Partial Digestion of Eukaryotic DNA for Use in Genomic Libraries: Pilot Reactions Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3978

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

Protocol is used to establish conditions for restriction enzyme digestion of eukaryotic genomic DNA that will generate fragments of a size appropriate for construction of genomic libraries. To construct a genomic library, the average length of the starting genomic DNA should be at least eight times the capacity of the vector. - [Read Partial Digestion of Eukaryotic DNA for Use in Genomic Libraries: Pilot Reactions Protocol]

Screening cDNA Libraries Constructed in Eukaryotic Expression Vectors Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4069

Category: DNA Protocols: cDNA Library Protocols: cDNA Library Construction Protocols

Two methods of screening pools of cloned cDNAs are discussed: transfection into mammalian cells and injection into Xenopus oocytes. - [Read Screening cDNA Libraries Constructed in Eukaryotic Expression Vectors Protocol]

Single-tube Coupled Transcription/Translation Reactions for Eukaryotic In Vitro - http://www.promega.com/tbs/tm045/tm045.pdf

Category: Cell Biology and Cell Culture: In Vitro Reconstitution Protocols

Information for protocol using single-tube, coupled transcription/translation reactions for eukaryotic in vitro translation. Includes information on: Translation Procedure; Positive Control Translation Reactions Using Luciferase; Cotranslational Processing Using Canine Pancreatic Microsomal Membranes; Post-Translational Analysis; Positive Control Luciferase Assays; Composition of Buffers and Solutions; Luciferase SP6/T7 Control DNAs - [Read Single-tube Coupled Transcription/Translation Reactions for Eukaryotic In Vitro]

Partial Digestion of Eukaryotic DNA for Use in Genomic Libraries: Preparative Reactions Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3979

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

The results of the pilot experiment (Partial Digestion of Eukaryotic DNA for Use in Genomic Libraries: Pilot Reactions) are used to establish conditions for partial digestion of eukaryotic DNA in the large-scale reactions described here. - [Read Partial Digestion of Eukaryotic DNA for Use in Genomic Libraries: Preparative Reactions Protocol]