Lab exam.docx - Vida Andoun Bisc 330 Lab-Friday 8:00 AM TA...

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Vida Andoun Bisc 330 Lab-Friday 8:00 AM TA: Jason Wang April 20, 2018 Identifying Unknown Samples Materials P10, P100, P1000 Micropipettor and tips Spectrophotometer Test Tubes and Racks UV/VIS Cuvettes DI H Methanol Bottles 12 mM O-NPG Solution Bradford Reagent (Coomassie Blue G-250, Phosphoric acid, and Methanol) 0.8 M Na 2 CO 3 - Dispenser Bottle Z Buffer 150 µL of three Unknown Samples Procedure Part A 1) Obtain 20 µL of each unknown sample to 3 test tubes and label them. 2) You will need to make 1:50 dilution of each sample for the Bradford and β-Gal assays and the total volume of 1 mL. 3) Add 980 µL of Z buffer to each test tube and vortex briefly, now you have three test tubes of 1:50 dilution. 4) Fill the table of 1:50 dilution. 5) Calculation C 1 V 1 = C 2 V 2 (1 × V 1 = (1/50) × (0.5 mL)) So, V 1 = 0.01 mL of cell extract. (if for measuring the absorbance at Part B, the dilution gives out of range absorbance try to make other dilutions, for example if 1:50 gives a reading 1.5 so it might give 0.75 at dilution 1:100. Keep in mind there is limited sample.) Part B 1. You need a new set of test tubes to perform Bradford assay. 2. Add 3.0 mL of Bradford reagent that is already pre-set.
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  • Spring '07
  • Petruska,Tower
  • Bradford protein assay, Coomassie, Bradford Assay, Bradford reagent

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