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Position：Dib ® SYBR qPCR SuperMix Plus
■ Product Description:
This product is a special reagent for Real Time PCR using SYBR Green I chimeric fluorescence method. In the premix, DNA polymerase,
The carefully optimized reaction Buffer, dNTPs, SYBR Green I and other reagents are premixed together, which is a 2× concentration premixed reagent, and can be used for solid
The preparation of PCR reaction solution is very simple and convenient.
This product uses a novel antibody modified HotStart Taq DNA polymerase, combined with a carefully optimized reaction Buffer, resulting in effective low inhibition
Non-specific amplification at temperature improves response specificity and amplification efficiency, enabling accurate quantification in a wider range.
■ Product features:
High specificity: Antibody modified HotStart Taq DNA polymerase is prepared by a novel process, which can perform PCR reaction without heat start-up.
Greatly improve the specificity of PCR amplification.
High efficiency: DiB carefully formulated RealTime PCR special 2×SuperMix for higher amplification efficiency and sensitivity.
Fast: the reagents necessary for PCR reaction are collected in one tube, and the reaction system can be prepared in a few minutes.
■ Suggestions:
Usage: mix evenly and gently upside down to avoid foaming, and apply after slightly centrifugation. The preparation and packaging of the reaction solution should be new (none
Contaminated) tips, microtubes, etc., to avoid contamination. The recommended reaction volume is 20-50 μ L.
Primer design: Generally, in order to increase sensitivity and amplification efficiency and inhibit non-specific response, the shorter the target sequence is, the better. Recommended design
Below 200bp.
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Save:
Store at -20℃ away from light. If you need to take it frequently in a period of time, it can be stored at 2~8℃ for 3 months. Avoid repeated freezing and thawing.
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Quality Assurance:
Purity test: all components were tested without endonuclease, exonuclease and nucleic acid residues.
■ Product packaging:
The component of product
A packaging
B packing
2 x dib ® SYBR qPCR SuperMix Plus
1 ml x 5
1 ml x 25
RNase Free Water
1 ml x 5
1 ml x 25
Reference Dye I (ROX I)
200 mu l
1ml
ROX Reference Dye II
200 mu l
1ml
ROX I calibrated real-time PCR apparatus includes:
ABI 5700/7000/7300/7700/7900/7900 HT/7900 HT Fast; ABI StepOne/StepOnePlus and other instruments.
Real Time PCR instruments calibrated with ROX II include:
ABI 7500/7500 Fast; ABI ViiA7; ABI Q5 Q6;
ABI Quant Studio 6/7 Flex; Stratagene MX4000/MX3500P/MX3000P and other instruments. Hong Kong ABC Biotech Co.,Limited
Room 502C,5th floor, Happy View Commercial Centre,
2-6 Garden Street,Mongkok, Kowloon, Hong Kong
Real Time PCR machines that do not require ROX calibration include:
Bio - Rad CFX96 provided/CFX 384 / iCycler iQ5 / / My iQ iQ5 MiniOpticon/an Opticon/an Opticon/Chromo4; Eppendorf
MasterCyclerrealplex/realplex 2 s.
Cepheid SmartCycler; Illumina Eco qPCR.
Roche Applied Science LightCycler 480;
Thermo Scientific PikoReal Cycler;
Qiagen/Corbett rotor-gene Q/ rotor-gene 3000/ rotor-gene 6000 and other instruments.
■ Operation method:
Common reaction system (20μ L) :
2 x DIB ® SYBR qPCR SuperMix Plus
10 mu l
The upstream primer
0.2 to 1.0 microns (end
Downstream primers
0.2 to 1.0 microns (end
The template
T
aq
1-2 mu l
ROX (depending on machine model)
0.4 u l
RNase Free Water
To 20 mu l
Common PCR cycles * :
* High specificity can be obtained by two-step method and high amplification rate can be obtained by three-step method.
Two-step amplification program:
95 ℃,
1 minute;
35-45 times cycle
95℃, 20 seconds;
60℃, 1 minute
Three-step PCR amplification procedure:
95 ℃,
1 minute;
95℃, 20 seconds;
35-45 times cycle
50-60℃, 20 seconds;
72 ℃,
30 seconds