Transfection procedures
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		Introduction
		We will express recombinant proteins in COS or HEK293 Cell  s Ð see table below for further details. COS (HEK293) cells will be provided at 50-70% confluency. We will use three different transfection procedures/reagents, DEAE dextran, calcium phosphate and Effectene™.
		Aims
		You will work individually, to perform your transfections Ð two people will work at the same time in the tissue culture facilities in the Wolfson building.
		Caution
		Class II GMOs Ð This practical involves the use of category II genetically modified organism -please follow instructions by demonstrators for safe handling and disposal.
		Timescale
		Cell  s must be split to an appropriate density, one or more days prior to transfection. Different transfection procedures take differing lengths of time - methodical planning is the key to success.
		Transfection strategy
		
		Effectene Transfection
		Materials required
		Condensation buffer (Buffer EC) pEGFP-N1
		Effectene Transfection Reagent pP2X2-EGFP
		Enhancer pGL3-BAS & pGL3-CON
		1x PBS  pcDNA3.1+






