Applied Biosystems 7500 - Before Starting the Installation; Setting Up the Computer; Selecting the Installation Method; Planning the Installation

Primer Extended on mRNA
5′
3′
Reverse
Primer
5′
Synthesis of 1st cDNA strand
cDNA
Oligo d(T) or random hexamer
3′
5′
cDNA
Applied Biosystems 7300/7500/7500 Fast Real-Time PCR System Relative Quantification Getting Started Guide 17
Notes
Chapter 3
3
Performing Reverse Transcription
Isolate total RNA
Adjust RNA
concentration
Convert
total RNA to cDNA
Performing
Reverse
Transcription
Intr
oduc
ti
on
and Exam
p
l
e
R
Q
Experimen
t
Desi
g
nin
g
an RQ
Experimen
t
P
r
i
m
e
r
E
xt
e
n
ded
o
n m
RNA
5
´
3
´
Revers
e
P
r
i
m
e
r
5
´
Sy
nthesis of 1st cDNA stran
d
c
DN
A
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
Seepage18
Seepage18
See page 19