CENTRIFUGATION3 - V.Woriax 8/09 BIO 3710 -Cell Biology-...

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V.Woriax 8/09 BIO 3710 --Cell Biology-- Differential Centrifugation of Tissue Homogenates Purpose: Use of mid-speed centrifuge; homogenization; techniques utilized when using biological samples PRECAUTIONS: All buffers used are non-toxic, aqueous solutions. Always wear eye protection when operating motor-driven equipment; always ensure that tubes are of equal volume and balanced opposite of each other before starting the centrifuge. Maintain sample tubes as cold as possible throughout the lab exercise to inhibit degradation of molecules. Items Needed: Beakers for homogenate (50 ml or larger) 0.25 M sucrose 12 ml Centrifugation tubes with caps (6/group) balances and weigh boats Test tube holders (beakers) chicken liver graduated cylinders tissue homogenizer spatulas (small red ones) scissors and forceps for cutting tissue pipettes/tips medicine droppers Perishable Needs: 0.25M Sucrose: MW = 342.3 Prepare 85.58 g sucrose/L Liver: Will need 6 g liver/group X 1 lb/453.6g = 0.0132 lb/group If 12 groups = 0.158 lbs total; 18 groups = 0.238 lbs total Procedure: NOTE: You will only need to retain one aliquot of each sample. KEEP SAMPLES ON ICE DURING THIS PROCEDURE AS MUCH AS POSSIBLE to reduce the action of degradative enzymes that may be released from broken lysosomes!!! At the end of this laboratory exercise, you will save 5 different biological samples. NOTE: Each step may need to be performed in duplicate depending upon the number of groups
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This note was uploaded on 03/30/2010 for the course BIO cell biolo taught by Professor ? during the Spring '10 term at University of North Carolina Wilmington.

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CENTRIFUGATION3 - V.Woriax 8/09 BIO 3710 -Cell Biology-...

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