Transcription of Protocols and tips in protein purification
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Protocols and tips in protein purification
Protocols and tips in protein purification or How to purify protein in one day Second edition 2018 Department of Molecular Biology & Biotechnology 2 3 Contents I. Introduction 7 II. General sequence of protein purification procedures Preparation of equipment and reagents Preparation and use of stock solutions Chromatography system Preparation of chromatographic columns Preparation of crude extract (cell free extract or soluble proteins fraction) Pre chromatographic steps Chromatographic steps Sequence of operations during IEC and HIC Ion exchange chromatography ()
ultra pure water to the top mark on the bottle. Worked example: to prepare 1 litre of buffer 50mM tris-HCl pH 8, 100 mM NaCl, 2mM EDTA: pour 50 ml of 1M tris-HCl, 20 ml of 5M NaCl and 10 ml of 0.2M EDTA in to a 1 litre bottle and add ultra pure water to 1 litre mark. Please note that dilution leads to a decrease in buffer pH.
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