Buffers - NOTE1 This year your TAs already prepared ALL the...

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Molecular Biology and Fermentation Buffers and solutions LB-agar x 4 500 mL Flask 5g LB 4g Agar 200mL DI water After filling water, cap the flask with a foam plug and wrap top with aluminum foil, autoclave TAE buffer for DNA electrophoresis 50X TAE (500mL) 121 g of Tris Base 28.05 mL of acetic acid 50mL of 0.5 M EDTA 1X TAE (1L) 20mL 50X TAE 980mL DI water Kanamycin = 10 mls 35g/ml and 0.22u filter sterilize Store 1ml aliquots in –20 Freezer 1M DTT – 5 mls (FW = 154.25g/mol) 0.77g in 5mls Store 1ml aliquots in –20 Freezer 1M IPTG – 10 mls (FW = 238.37 g/mol) 2.384g in 10mls and 0.22u filter sterilize Store 1ml aliquots in –20 Freezer 1M NaOH – 300 mls In a beaker with a stir bar add 200ml DI water Add 12g NaOH Be careful, the solution releases heat Dissolve Adjust final volume to 300mls Place a label and your group number on the jar 1M HCl – 300mls (stock HCl = 12M) Warning : Always add acid to water 275mls DI water + 25mls 12M HCl Downstream processing and purification
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