solution and dilution 1) You have a 50 mM Tris-HCl stock solution and a 125 mM E
ID: 175888 • Letter: S
Question
solution and dilution
1) You have a 50 mM Tris-HCl stock solution and a 125 mM EDTA stock solution. How will you use these stock solutions to make 500 ml of TE Buffer? (TE Buffer has 10 mM TrisHCl and 1 mM EDTA).
2) You have transformed your bacterial cells with the plasmid pLux and now you would like to isolate this DNA from the cells. The protocol requires that you resuspend your pellet of bacterial cells in Solution I containing 50mM Glucose, 25mM Tris and 10mM EDTA. You have the following stock solutions 100mM Glucose; 1M Tris; 0.5M EDTA. Describe how you would prepare 100ml of Solution I.
Explanation / Answer
Answer 1.
TE Buffer has 10 mM Tris-HCl and 1 mM EDTA. You have to prepare 500 ml of the buffer.
Tris-HCl stock is 50mM. Now, use the formula,
N1 * V1 (stock) = N2 * V2 (working solution); final working solution should have 10mM Tris-HCl.
50* V1 = 10* 500
V1 (Tris-HCl) = 100ml
EDTA stock is 125mM. Now, use the formula,
N1 * V1 (stock) = N2 * V2 (working solution); final TE buffer should have 1mM EDTA.
125* V1 = 1* 500
V1 (EDTA) = 4ml
So, to prepare 500 ml TE buffer, add 4ml EDTA and 100ml Tris-HCl stock in to 396ml distilled water.
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