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| ==Protocol==
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| ===Day 1===
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| ====Equipment====
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| *Fluorometer
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| *Plate x1
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| *Plate Centrifuge
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| *6x1.5ml tubes
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| *Gilson Pipettes
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| '''Reagents and Chemicals'''
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| #Remove pure sample of recombinant GFP protein of .....M from storage
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| #Remove .... of our cell extract solution from storage solution and place in a 1.5ml tube and put to the side.
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| #Now we need to prepare 100μl of the commercial E.coli and T7 cell extract solution. First we need to prepare complete amino acid mixture for both extract solutions: Add the 7.5μl volume of two amino acid minus mixtures for both kits''<br>Now prepare the commercial cell extract:<br>
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| #Take 3x1.5ml tube and add 5µl of the ''E.coli complete amino acid mixture''
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| #Take 3x1.5ml tube and add 5µl of the ''T7 complete amino acid mixture''
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| #To each 1.5ml tube add 20µl of ''S30 Premix Without Amino Acid''
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| #Add 15µl of ''S30 Extract Circular''
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| #Add 55µl of ''nuclease-Free Water'' to make volume upto 100μl
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| '''Prepare GFP serial Dilations'''
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| #The commercial S30 extracts are claimed to produce around 150-200ng of protein. We need to cover this range within our calibraation curve.
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| #Prepare suitable dilutions given a suitable range:
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| #Remember need to consider the concentration of GFP in total volume of cell extract
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| '''Loading Plate'''
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| #Follow the schematic for loading the plate, begin by adding the relevant cell extracts to the correct wells.
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| #Then add the correct dilution of GFP to the corrrect well.
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| #Place place in the fluorometer and measure the intial fluorescence, try to minimise the time between addition of the GFP and reading the plate.<br>
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| '''Schematic'''
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