====== General ====== 0.1M MgCl2 * 0.813g MgCl2 (MW 203.31) * 40mL H2O * filter 1M Tris * 121g Tris base * 900mL H2O * filter 1M DTT * 1.543g DTT (MW 154.3) * 10mL H2O APS, 10% w/v * 5g Ammonium Persulfate (APS) * 50mL H2O 0.25M Spermidine trihydrochloride * 0.636g Sperm (MW 254.63) * 10mL H2O * filter ====== RNA/DNA Related ====== 100mM NTPs ^rATP^rGTP^rCTP^UTP^Mix^ |276mg rATP (MW 551.1)|262mg rGTP (MW 523.2)|264mg rCTP (MW 527.1)|242mg UTP (MW 484.1)|rATP + rGTP + rCTP + UTP| |5mL H2O|5mL H2O|5mL H2O|5mL H2O|5mL H2O| |Add 50mM (500mM?) Tris Base to adjust pH to 7 (by pH paper) and filter||||| RNA Loading buffer, 50 mL * 13 mg Bromphenol Blue * 10 mg Xylene cyanol * 930 mg EDTA * solve in 50 mL Formamide, heat briefly DNA Loading buffer, 10 mL * 25 mg Bromphenol Blue * 25 mg Xylene cyanol * 10mL 30% glycerol Agarose Gel * 2g Agarose * 100mL 1X TAE * microwave until boiling * wait until warm and add 2.5uL EtBr 15% Polyacrylamide, 8M Urea, 1X TBE, 2L * 960g Urea * 400mL 5X TBE * 750mL 40% Polyacrylamide * Stir over gentle heat Crush & Soak Buffer * Tris 20mM * Sodium acetate 300mM * EDTA 1mM 8M Urea, 1X TBE, 2L * 960g Urea * 400mL 5X TBE * 1L DEPC H2O