Prepare 100ml of homogenization buffer. Calculate how to do this with (c1V1=c2V2
ID: 980694 • Letter: P
Question
Prepare 100ml of homogenization buffer. Calculate how to do this with (c1V1=c2V2) where C is concentration and V is volume.
Initial Stock solutions:---------------------- Final Buffer:
50mM TrisCl, pH8.6 ---------------------------10mM TrisCl, pH 8.6
100mM PMSF-----------------------------------1mM PMSF
20mM EDTA--------------------------------------1mM EDTA
20 uM pepstatin A -------------------------------0.2 uM pepstatin A
7.1 M 2-mercaptoethanol ----------------------1 mM 2-mercaptoethanol
Please show your work so I can reason through this! Thanks
Explanation / Answer
C1V1 = C2V2
C1 – Concentration of Initial stock solution
V1 – Volume of Initial stock solution to be taken for preparation of final solution
C2 – Concentration of Final solution
V2 – Volume of Final solution
50mM Tris Hcl
C1 – 50mM
V1 – ?
C2 – 10mM
V2 – 100ml
So V1 = (10 x 100)/50
V1 = 20ml
100mM PMSF
C1 – 100mM
V1 – ?
C2 – 1mM
V2 – 100ml
So V1 = (1 x 100)/100
V1 = 1ml
20mM EDTA
C1 – 20mM
V1 – ?
C2 – 1mM
V2 – 100ml
So V1 = (1 x 100)/20
V1 = 5ml
20µM Pepstatin A
C1 – 20µM
V1 – ?
C2 – 0.2µM
V2 – 100ml
So V1 = (0.2 x 100)/20
V1 = 1ml
7 M -Mercaptoethanol
C1 – 7 M
V1 – ?
C2 – 1mM
V2 – 100ml
So V1 = ((1 x 100)/7)/1000
V1 = 0.014ml
1000 is divided for converting Molar into Millimolar
Mix the above said volumes in a 100ml measuring cylinder or volumetric flask and make upto 100ml with the solvent that is going to dissolve