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CALCULOS

REACTIV
O
CONCENTRACI
N
STOCK
CONCENTRA
CIN FINAL
CLCULOS
Buffer
PCR
10X 1X C
i

*
V
i
= C
f *
V
f
1X
*
20 mL = 10 X
*
V
f
2 mL = V
f
dNTPs 2.5 mM 0,5 mM C
i

*
V
i
= C
f *
V
f
0,5 mM
*
20 mL = 2.5 mM
*
V
f
4 mL = V
f
MgCl
2
50 mM 2 mM C
i

*
V
i
= C
f *
V
f
2 mM
*
20 mL = 50 mM
*
V
f
0,8 mL = V
f
Prier 10 mM 0,5 mM C
i

*
V
i
= C
f *
V
f
0,5 mM
*
20 mL = 10 mM
*
V
f
1 mL = V
f
T!"#
P$lier!
s!
5 m/ mL 0,125 m/ mL C
i

*
V
i
= C
f *
V
f
20 mL
*
0,125 m/ mL = 5 m/
mL
*
V
f

0,5 mL = V
f
Los resultaos a!teriores e"e! ser multi#li$aos #or 5, %a &ue se
reali'ara! 4 e!sa%os (u!o #or $asa i!te)ra!te* mas el $o!trol+
a. ,uffer -C.+ V
f
= 2 mL
*
5 = 10 mL
". /0-1s+ V
f
= 4 mL
*
5 = 20 mL
$. M)CL
2
+ V
f
= 0,8 mL
*
5 = 4 mL
. -rimer+ V
f
= 1 mL
*
5 = 5 mL
e. 0a&. -olimerasa+ V
f
= 0,5 mL
*
5 = 2.5 mL
La suma e toos los rea$ti2os so!+ 41,5 mL a lo $ual se le e"e! a)re)ar
los 5 mL e A3/ &ue e"emos utili'ar. 4! total te!emos 45,5 mL.
4l 2olume total es e 100 mL, #or lo e e"emos restar el 2olume! total ara
sa"er $ua!to ser6 lo e a)ua &ue o$u#aremos+
100 mL 7 45,5 mL = 58,5 mL (9
2
O*