Question

1) Write out your calculations for preparing 2% SDS lysis buffer (use M1V1=M2V2)

In an Eppendorf tube, prepare 1 mL of 2% SDS lysis buffer (M1V1 = M2V2) 2% SDS lysis buffer 1 mL-1 2% SDS (stock = 20% 50 mM Tris-HCl, pH 8 (stock = 1 M) MV, = M,V2 10 mM EDTA (stock = 0.5 M) Bring up to Vf of 1 mL with diH20 Store at room temperature

2) Based on these calculations, how many micrograms of protein are in 10 microliters of each sample?

Answer 1

(1) For dis sps lysis buffer using mv = M₂. Ve where, on, a nitial concentration V = quitial volume M2 = Final concentration V2 = Final volume here, for 2% SDS M = 26% M2 = 2% V=? , Vr = 1 .. 20% XV = 77. xlmy - 10 m = 0.1 me for Tris-Hol M = M, M₂ = 50mm = Soxlor) M = 0.05m V = ? , Vr 1 m 4 x = 0.05th x 1m. i = 0.05 me for EDTA 0.01 M = 0.5m, M2= 10mm= = ? , V2 = 1 in 0.54 x = 0.014 x 1m. .: v= 0.01xl am = 0.02m To make lysis buffer of 2% 2% SDS needed = olm This Hol = 0.05m EDTA = 0.02 mi di ho needed = 16.1 +0.05 +002) and = 0.83 del to make total volume of Ime (2) To answer this protein concenration needed.