Hi, I really need up fiuring out how to do this. I\'m doing a 1:10 ratio dilutio
ID: 49992 • Letter: H
Question
Hi, I really need up fiuring out how to do this. I'm doing a 1:10 ratio dilution. I know it's regular chem. lab but I can't remember. Equation and help please. Need by tonight.
Use 0.2 ml PCR tubes for these reactions. The thin wall facilitates heat transfer. Be careful to not crush the tubes between your fingers. There is not much writing surface on these tubes so it is best to create a log in your notebook and then number the tubes accordingly.
Reagent
Stock Conc
Final Conc
Dilution
Volume per Sample
Volume per Master Mix (N+1)
Template DNA
5 ul
5X GoTaq Flexi Buf
5X
1X
dNTP Mixture
10 mM
0.2 mM
MgCl2
25 mM
2.5 mM
D1S80 Fwd Primer
10 uM
0.5 uM
D1S80 Reverse Primer
10 uM
0.5 uM
GoTaq Hot Start Polymerase
5U/ul
1.25U
MGW (Molecular Grade Water)
Reagent
Stock Conc
Final Conc
Dilution
Volume per Sample
Volume per Master Mix (N+1)
Template DNA
5 ul
5X GoTaq Flexi Buf
5X
1X
dNTP Mixture
10 mM
0.2 mM
MgCl2
25 mM
2.5 mM
D1S80 Fwd Primer
10 uM
0.5 uM
D1S80 Reverse Primer
10 uM
0.5 uM
GoTaq Hot Start Polymerase
5U/ul
1.25U
MGW (Molecular Grade Water)
Explanation / Answer
10 mM dNTPs (2.5mM each dT, dA, dC, dGTP) - 4.0l -- volume per master mix (N+1) = 20l
Gene Specific Primer mix -1.0l -- volume per master mix (N+1) = 5l.
10X PCR Buffer (includes MgCl2) 5.0 l -- volume per master mix (N+1) = 25l .
Taq polymerase (this is in the 0.2 ml tube already ) 0.25l
dH2O -- 38.75l; volume per master mix (N+1) = 193.75l.
The primer mix contains both PCR primers at concentrations of 10 pmoles/l each. The10X PCR buffer will give a final reaction concentration of 50 mM Tris-HCL pH 9.1, 16mM ammonium sulfate, 3.5 mM MgCl2and 150g/ml BSA.
Related Questions
drjack9650@gmail.com
Navigate
Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.