ShineProbe®
Real Time qPCR
MasterMix Kits Ⅰ、Description
ShineProbe
is a 2X convenient premix reagent, specially designed
for real time PCR by using various detection probes (eg.
TaqMan, Molecular Beacon, etc.) ，but
not suitable for sybr green.The
master mix contains all reagents needed for qPCR. Only
template ,Probe,and PCR primers need to be added by the
user.The
ROX dye is not provided,
For
most real-time instruments ROX passive reference dye is
not required
.This product combines the high performance of Taq,
which is an enzyme for hot start PCR utilizing Taq
antibody, with a newly developed buffer which provides
superior specificity, increased amplification efficiency
and high aptitude for high-speed real time PCR.
Accordingly, a successful real time PCR is promised with
high sensitivity, wide dynamic range and accurate
quantification.
The reaction chemistry of ShineProbe qPCR Kit is
applicable to most real-time PCR instruments, e.g. from
Applied Biosystems, Bio-Rad Laboratories, Corbett
Research, Roche,and Stratagene. Ⅱ、Component ZK0071350reactions
X 50ul eachHotstart fluo-PCR mix
(2
X conc.)
ZK00714100
reactionsX
50ul each Hotstart
fluo-PCR mix (2
X conc.) Ⅲ、Shipping
and Storage
The
ShineProbe qPCR Kit is shipped in dry ice or ice bag.
Upon arrival, store all kit components
at -20°C. When using the 2x master mix, the leftover
thawed mix can be refrozen
and stored at -20 °C without affecting the performance
of the kit. The kit is stable for six months from the
date of packaging when stored and handled properly.
Ⅳ、Reaction
Setup
Components
Volume(μl)
Final
conc.
Comments
Hotstart
Fluo-PCR mix
25
1
X
Mix
thoroughly
Sense
primer(25pmol/ul)
1
0.3~1uM
Titrate
from 0.3 to 1 µM if necessary.
Anti-sense
primer(25pmol/ul)
1
0.3~1uM
Titrate
from 0.3 to 1 µM if necessary.
Probe(25pmol/ul)
0.5
0.15~0.5uM
cDNA(ul)
2
10~100ng
ddH2O
(ul)
20.5
-
Add
water to fill up to the final
reaction volum
Total
Volume(ul)
50
-
Ⅴ、Cycling
Protocol
1、Protocol
using LightCycler with Taqman probe
93℃
2min→93℃5
sec→60℃30
sec
40cyclers
2、Protocol
using LightCycler with Molecular Beacon probe
93℃
2min→93℃5
sec→60℃20
sec→72℃20
sec
40cyclers
3、Protocol
using other instruments,
e.g. from Applied Biosystems, Bio-Rad Laboratories,
Corbett Research, ,and Stratagene.
with Taqman probe
94℃
4min→94℃
30sec→60℃
60sec
40cyclers
4、Protocol
using other instruments,
e.g. from Applied Biosystems, Bio-Rad Laboratories,
Corbett Research, ,and Stratagene.
with Molecular Beacon probe 94℃
4min→94℃
30sec→60℃
30sec→72℃30sec 40cyclers
