please show your work A student has a let-23 gene with concentration 79ul Now he
ID: 203410 • Letter: P
Question
please show your work
A student has a let-23 gene with concentration 79ul Now he wants to know the amount needed per 30 ul reactions. Fill in the boxes for the appropriate amounts needed for the reactions Component Final concentration Amount needed per 30 ul reaction 20 nglu plasmid DNA 10X EcoRI Buffer 1 U/ul EcoR mol grade water EcoRI 0.2 U/ul total volume 30 ul Component Final concentration Amount needed per 30 ul reaction plasmid DNA 10X Buffer 1 10X BSA mol grade water 20 ng/ul Sacl 1X total volume 30 ulExplanation / Answer
Plasmid DNA stock concentration = 79ng/ul
EcoR1:
Plasmid DNA
Plasmid DNA stock concentration = 79ng/ul
Required final plasmid DNA concentration = 20ng/ul
Final volume = 30ul
M1V1 (Concentrated) = M2V2 (Diluted)
79ng/ul * xul = 20ng/ul*30ul
Xul = 20ng/ul*30ul/79ng/ul
= 7.594ul
We have to take 7.594ul from 79ng/ul stock plasmid DNA to get 20ng/ul for 30ul reaction mixture
10X EcoR1 buffer:
Stock buffer concentration = 10X
Required final buffer concentration = 1X
M1V1 (Concentrated) = M2V2 (Diluted)
10X * xul = 1X*30ul
xul = 1X*30ul/10X
= 3ul
We have to take 3ul from 10X EcoR1 buffer stock to get 1x for 30ul reaction mixture
EcoR1 enzyme:
EcoR1 enzyme stock concentration = 1u/ul
Required final plasmid DNA concentration =0.2u/ul
Final volume =30ul
M1V1 (Concentrated) = M2V2 (Diluted)
1u/ul * xul = 0.2u/ul*30ul
Xul = 0.2u/ul*30ul/1u/ul
= 6ul
We have to take 6ul from EcoR1 enzyme stock to get 0.2u/ul for 30ul reaction mixture
For to 30ul Reaction mixture:
S.No.
Component
Final concentration
Amount needed for 30ul reaction
1
Plasmid DNA stock (79ng/ul)
20ng/ul
7.594ul
2
10X EcoR1 buffer
1X
3ul
3
EcoR1 enzyme stock (1u/ul)
0.2u/ul
6ul
4
Mol grade H2O
-
13.406ul
Total volume 30ul
SacI
Plasmid DNA
Plasmid DNA stock concentration = 79ng/ul
Required final plasmid DNA concentration =20ng/ul
Final volume =30ul
M1V1 (Concentrated) = M2V2 (Diluted)
79ng/ul * xul = 20ng/ul*30ul
Xul = 20ng/ul*30ul/79ng/ul
= 7.594ul
We have to take 7.594ul from 79ng/ul stock plasmid DNA to get 20ng/ul for 30ul reaction mixture
10X buffer1:
Stock buffer concentration = 10X
Required final buffer concentration = 1X
M1V1 (Concentrated) = M2V2 (Diluted)
10X * xul = 1X*30ul
xul = 1X*30ul/10X
= 3ul
We have to take 3ul from 10X buffer stock to get 1x for 30ul reaction
10X BSA:
Stock BSA concentration = 10X
Required final buffer concentration = 1X
M1V1 (Concentrated) = M2V2 (Diluted)
10X * xul = 1X*30ul
xul = 1X*30ul/10X
= 3ul
We have to take 3ul from 10X BSA stock to get 1x for 30ul reaction
For to 30ul Reaction mixture:
S.No.
Component
Final concentration
Amount needed for 30ul reaction
1
Plasmid DNA stock (79ng/ul)
20ng/ul
7.594ul
2
10X buffer 1
1X
3ul
3
10X BCA
1X
3ul
4
Mol grade H2O
-
16.406ul
Total volume 30ul
S.No.
Component
Final concentration
Amount needed for 30ul reaction
1
Plasmid DNA stock (79ng/ul)
20ng/ul
7.594ul
2
10X EcoR1 buffer
1X
3ul
3
EcoR1 enzyme stock (1u/ul)
0.2u/ul
6ul
4
Mol grade H2O
-
13.406ul
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