Concentration in 1x buffer Concentration in 10x Stock concentrations Vol of stoc
ID: 190616 • Letter: C
Question
Concentration in 1x buffer Concentration in 10x Stock concentrations Vol of stock solution to make buffer (rom previous page) 10x buffer (ml) 50mM Tris HCl pH 8.0 IM 10mM MgC 100mM NaCI 100 lg/ml BSA H,O to final vol IM 5M 10 mg/ml 2. A very common reaction used in molecular biology laboratories is the cutting of DNA molecules with restriction enzymes. Plasmid DNA: a small circular DNA molecule. We'll learn more about these later Restriction enzyme: an enzyme that cuts DNA at specific sequences. They're supplied in concentrations of "units", or U, of enzymatic activity. We'l learn more about them later, too amenrations of"units"', or U.o Using the 10X buffer we made above, DNA and an enzyme, try setting up the reaction describeExplanation / Answer
1) Stock plasmid DNA concentration =50ug/ml
Required plasmid DNA concentration (final conentration) =0.5ug in 50ul of reaction
M1V1 (Concentrated) = M2V2 (Diluted)
50ug * xul (?) = 0.5ug (Final concetion we need) * 50ul (Reaction final volume)
xul = 0.5ug*50ul/ 50ug
=0.5ul
So we have to take 0.5ul of stock plasmid DNA for 50ul reaction mixture
2) Stock buffer concentration =10X
Required final reaction buffer concentration =1X
10X * xul = 1X*50ul
xul = 1X*50ul/10X
= 5ul
We have to take 5ul from stock to get 1x for 50ul reaction mixture
3) Stock enzyme concentration =2units/ml
Required final enzyme concentration =1 unit
2units * xul = 1 unit*50ul
xul = 1 unit *50ul/2units
= 25ul
We have to take 25ul from stock enzyme to get 1 unit for 50ul reaction mixture
For to 50ul Reaction mixture:
10 reaction buffer = 5ul
50ug Stock DNA =0.5ul
2units stock enzyme =25ul
H2O =19.5ul
Total volume =50 ul
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