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Great scientific discoveries have been made by men seeking to verify quite erroneous theories about the nature of things. ~Aldous Huxley, "Wordsworth in the Tropics"
Protocols for transfection, lipofection, transfecting RNA, DNA, proteins and RNAi into cells.
Calcium Phosphate Transfection Protocols (5)Electroporation Sonoporation Protocols (15) | Stable Transfection Protocols (6)Transient Transfection Protocols (7) |
| COS Cell Transfection Protocol - http://genetics.med.harvard.edu/~cepko/protocol/mike/C5.html COS cell transfection protocol. Includes solutions: DEAE Dextran and 1000X Chloroquine - [Read COS Cell Transfection Protocol] |
| Dlgh1 Knockdown in Primary OT-1 T Lymphocytes: Transfection Protocol - http://www.natureprotocols.com/2007/01/29/dlgh1_knockdown_in_primary_ot1.php Transfection protocol used in the Dlgh1 knockdown in primary OT-1 T lymphocytes. - [Read Dlgh1 Knockdown in Primary OT-1 T Lymphocytes: Transfection Protocol] |
| DNA Transfection Mediated by Lipofection Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3870 Protocol should be viewed as a starting point for systematic optimization of transfection mediated by lipofecting agents. Once a positive signal has been obtained from a transfected plasmid carrying a standard reporter gene, optimal conditions for transfection can be established by systematic variation of parameters such as the initial cell density, the amount and purity of DNA, the media and serum, and the time of exposure of the cells to the cationic-lipid-DNA complex. - [Read DNA Transfection Mediated by Lipofection Protocol] |
| DNA Transfection Using Polybrene Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3875 Polybrene and DMSO can be used to achieve stable transformation of several types of cells by plasmid DNA. The yield of transformants is up to 15-fold greater with Polybrene than with calcium phosphate-DNA coprecipitation. However, there is no difference between the two methods in the efficiency of transformation of cells by high-molecular-weight DNA. - [Read DNA Transfection Using Polybrene Protocol] |
| Optimizing Electrotransfection of Mammalian Cells In Vitro Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/1/pdb.prot4449 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] |
| Transfecting and Plating RAW 264.7 Cells with Lipofectamine 2000 Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000182.pdf?pid=PP00000182 The following procedure is for simultaneous transfection and plating of RAW 264.7 cells. This protocol results in approximately 50% to 70% cell viability, and of those viable cells, 20% to 40% are transfected when using pEYFP-N1 from Clontech. Include: Procedure for Splitting Cells before Transfection; Procedure for Preparing Lipofectamine 2000 and DNA; Preparation of RAW 264.7 Cells for Transfection. - [Read Transfecting and Plating RAW 264.7 Cells with Lipofectamine 2000 Protocol] |
| Transfection Mediated by DEAE-Dextran: High-efficiency Method Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3876 DEAE-dextran is generally used to obtain a burst of transient expression of cloned genes after transfection of mammalian cells. Many variants of the technique have been described, all of which seek to maximize the uptake of DNA and to minimize the cytotoxic effects of DEAE-dextran. In this protocol cells are exposed briefly to a high concentration of DEAE-dextran-DNA and then to chloroquine diphosphate, which is a facilitator of transfection. - [Read Transfection Mediated by DEAE-Dextran: High-efficiency Method Protocol] |
| Transfection of IMEC Cells Using SuperFect Transfection Reagent Protocol - http://www.flemingtonlab.com/Protocols/SuperfectionofIMECcells.pdf Transfection of IMEC cells using SuperFect Transfection Reagent protocol. Protocol includes experimental design considerations. - [Read Transfection of IMEC Cells Using SuperFect Transfection Reagent Protocol] |
| Transfection of Mammalian Cells with siRNA Duplexes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4343 Protocol describes a method for the delivery of siRNAs into mammalian cells in the absence of reporter plasmids. This is best achieved with transfection reagents developed for the delivery of antisense oligodeoxynucleotides. The quantities of reagents given below are calculated for the transfection of one well of a 24-well plate. - [Read Transfection of Mammalian Cells with siRNA Duplexes Protocol] |