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: NexQ-6000
Package : 1 ml
NEXpro qRT-PCR 2X Master Mix (Probe) is an optimized ready-touse solution for real-time quantitative RT-PCR assays, Using Taqman
probe etc..
It comprises all the components necessary to perform qRT-PCR:
MMLV (RNAse H-),Hotstart Taq DNA Polymerase, ultrapure dNTPs,
MgCl2. The user simply needs to add water, template,probe and
primers. An enhanced buffer allows for RT reaction temperatures up
to 50C. This can improve detection of more difficult targets as
higher RT temperatures reduce nonspecific priming and facilitate
melting of RNA secondary structures.
The kit includes the components necessary for performing RT-PCR
amplification, and have been successfully used to DNA amplify and
detect a variety of RNA targets.
Recommended Protocol
10 l
25 l
1X
0.2~2.0 l
0.5~5.0 l
0.1~1.0 M
0.2~2.0 l
0.5~5.0 l
0.1~1.0 M
Variable
Variable
Variable
1 l
1 l
as needed
up to 20 l
up to 50 l
NA
qRT-PCR Master(Probe)
Kit Contents
Applications
Fluorescence Probe
Real-time RT-PCR
Gene expression profiling
Gene knockdown verification
Array validation
Template RNA
Water, RNase-Free
Quality Control
Storage Conditions
Note
PCR Conditions
ROX reference dye is not included in this kit and may be added to
compensate for non-PCR related variations in fluorescence. Addition
of the reference dye is optional. Optimizing the ROX dye concentration within the qRT-PCR reaction is an important aspect of setup.
Too much ROX in the qRT-PCR reaction will reduce background but
also makes a low target signal difficult to distinguish from
background.
Step
Temp (C)
Time
Cycle
Reverse Transcription
45-50
10-30 min.
Initial Denaturation
95
10 min.
Denature
95
20 sec.
Anneal
50 ~ 65
40 sec.
Extend
72
60 sec./kb
Final Extension
72
5 min.
25~ 40
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