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When preparing samples for SDS-PAGE, what is the purpose of boiling the samples at 95°C?

Basic Clinical Laboratory Techniques 6E

6th Edition

ISBN:9781133893943

Author:ESTRIDGE

Publisher:ESTRIDGE

Chapter1: The Clinical Laboratory

Section1.8: Laboratory Math And Reagent Preparation

Problem 14RQ

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How would you make two-fold serial dilutions such that the last tube is a 1:32 dilution of the original, concentrated material? Assume that you need to have at least 500 µl of each dilution, and you should be able to perform the dilutions in microfuge tubes with a maximum capacity of 1.5 ml.
Design a serial dilution of a sample to achieve a A) Final dilution of 10-4 using 9.0 mL blanks B) Final dilution of 10-6 using 99 mL blanks C) Final dilution of 10-6 using 9.0 mL blanks
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My question is: Could you explain steps? (etc. why do we heat or add chemicals like nacl or ammonium sulphate?) 1)Casein Isolation from MilkExperimental Procedure* Put 100 ml of milk in a 500 ml beaker and heat it to 40 0C. * Also heat 100 ml acetate buffer and slowly add it to the beaker with milk while stirring. * The final pH of the mixture should be 4.8. * Cool the suspension to room temperature and wait 5 more minutes before filtering. * Wash the precipitate several times with small volumes of water and then suspend it in approximately 30 ml of ethanol.Filter the suspension in the buchner funnel and wash the precipitate a second time with a mixture of equal proportions of ethanol and ether. * Finally, wash the precipitate on filter paper with 50 ml of ether and soak and dry. * Make the ether evaporate by spreading the powder. * Weigh the casein and calculate the percentage yield of the protein. * Compare with theoretical yield of 3.5 g per 100 ml milk. 2)Purification of Egg…
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