Applied Biosystems 7500 - Designing an RQ Experiment

Applied Biosystems 7300/7500/7500 Fast Real-Time PCR System Relative Quantification Getting Started Guide 7
Notes
Chapter 2
2
Designing an RQ Experiment
Select the PCR method
Specify the components of an
RQ experiment
Select one-step or
two-step RT-PCR
Select the chemistry
Choose the probes
and primers
Designing
an RQ
Experiment
Intr
oduc
ti
on
and Exam
p
l
e
R
Q
Experimen
t
Performin
g
R
evers
e
T
ranscription
P
r
i
m
e
r
E
xt
e
n
ded
o
n m
RNA
5
´
3
´
Revers
e
Primer
5
´
Sy
nthesis of 1st cDNA stran
d
cDNA
O
ligo d(T) or random hexame
r
3
´
5
´
cDNA
Performin
g
an R
Q
S
tudy
OR
Generatin
g
D
a
t
a
fr
o
m
R
Q
Plates
S
tandard
STANDARD
TANDAR
Generatin
g
Da
t
a
fr
om
RQ
Plate
s
Fas
t
Seepage8
Seepage9
Seepage11
Seepage12
See page 14