1 Calculate and enter in Table below how much BSA stock solution (mL) and buffer solution (mL) are required to generate 5 serial dilutions with DF 1 2 2 Calculate and enter in Table below how much in volume (mL) of undiluted samples (A to E) and buffer solution (mL) are required to generate 1 mL (1000 mL) of A1 to E1 solutions by parallel dilution (1 st dilution) with DF 1 1 50 3 Continue from the 1 st parallel diluted sample, calculate, and enter in Table below how much in volume (mL) of A1 to D1 to generate 0 5 mL (500 mL) of solutions A2 to D2 by make one serial dilution (2 nd dilution) with DF2 1 10 Sample ID volume (ul) Final volume left in the tube (ul) volume (ul) of buffer solution Final concentration of BSA standard solution (mg mL) BSA stock solution Std 6 1000 0 0 10 500 Std 5 500 500 0 05 Std 4 500 Std 3 500 Std 2 500 Std 1 Keep 500 500 (Discard the rest) Blank 0 500 500 To make 1000 ul of 1st parallel dilution (DF1 1 50), take UL Sample ID To make 500 ul of 2nd serial dilution (DF2 1 10) from the previous dilution take UL Sample ID Sample ID Final DF Final (DF1 x DF2) volume in the tube (ul) Undiluted diluent diluent 1st diluted sample sample A A1 A2 500 B B1 B2 500 C C1 C2 500 D D1 D2 500 E E1 1 50 Keep 500 (Discard the rest)

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1. Calculate and enter in Table below how much BSA stock solution (mL) and buffer solution (mL) are required to generate 5 serial dilutions with

1. Calculate and enter in Table below how much BSA stock solution (mL) and buffer solution (mL) are required to generate 5 serial dilutions with DF = 1/2.

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2. Calculate and enter in Table below how much in volume (mL) of undiluted samples (A to E) and buffer solution (mL) are required to generate 1 mL (1000 mL) of A1 to E1 solutions by parallel dilution (1^st dilution) with DF₁ = 1/50

3. Continue from the 1^st parallel diluted sample, calculate, and enter in Table below how much in volume (mL) of A1 to D1 to generate 0.5 mL (500 mL) of solutions A2 to D2 by make one serial dilution (2^nd dilution) with DF2= 1/10.

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Sample ID volume (ul) Final volume left in the tube (ul) volume (ul) of buffer solution Final concentration of BSA standard solution (mg/mL) BSA stock solution Std. 6 1000 0 0.10 500 Std. 5 500 500 0.05 Std. 4 500 Std. 3 500 Std. 2 500 Std. 1 Keep 500 500 (Discard the rest) Blank 0 500 500 To make 1000 ul of 1st parallel dilution (DF1 = 1/50), take UL Sample ID To make 500 ul of 2nd serial dilution (DF2 =1/10) from the previous dilution take UL Sample ID Sample ID = Final DF Final (DF1 x DF2) volume in the tube (ul) Undiluted diluent diluent - 1st diluted sample sample A A1 A2 500 B B1 B2 500 C C1 C2 500 D D1 D2 500 E E1 1/50 Keep 500 (Discard the rest)