BioDiamond 2X qPCR Mix With 4x1.25ml 即時PCR混合酵素
Cat No. DMDRT001
Pack size: 4x 1.25 ml
qPCR Instruments:
BioRad CFX96, Roche LightCycler 480, MJ Research Opticon and Opticon 2, MJ Research Chromo 4, Corbett Rotor-gene 600,3000 and Eppendorf Realplex 2Product Application.
Product Description
2x qPCR Master mix is designed for quantitative real-time analysis of DNA samples.
•Strong signals and high sensitivity due to fluorescent dye.
•High specificity - no primer dimers, no NTC signal.
•Optimized 2x qPCR Master mixes for different real-time PCR instruments. Master mix formulations optimized for different machines. 2x qPCR Master mix is ideally suited for:
•Gene expression analysis
•Microarray validation
•Viral load determination qRT-PCR
Kit Components
2x qPCR Master mix is a 2X mix of dNTPs, Hotstart Taq polymerase, MgCl2, fluorescent detection dye, reference dye(optional), and proprietary buffer components.
Storage
long time: -20°C
short time: 4°C
Recommended Protocol
Thaw 2x qPCR Master mix, template DNA, RNase-free water and primer on ice. Mix each solution well. Prior to the experiment, it is prudent to carefully optimize experiment Conditions and to include controls at every stage. See pre-protocol considerations for details. This standard protocol applies to a single reaction where only template, primers and water need to be added to the 2X qPCR Master mix. For multiple reactions, scale-up volume of reaction components proportionally. All reagents should be thawed on ice, gently mixed and briefly centrifuged before use.
1.Thaw reagents at room temperature. Mix thoroughly and then place on ice immediately after thawing.
2. Assemble reaction tubes on ice.
3. Prepare a reaction Master mix using the following:
Pack size: 4x 1.25 ml
qPCR Instruments:
BioRad CFX96, Roche LightCycler 480, MJ Research Opticon and Opticon 2, MJ Research Chromo 4, Corbett Rotor-gene 600,3000 and Eppendorf Realplex 2Product Application.
Product Description
2x qPCR Master mix is designed for quantitative real-time analysis of DNA samples.
•Strong signals and high sensitivity due to fluorescent dye.
•High specificity - no primer dimers, no NTC signal.
•Optimized 2x qPCR Master mixes for different real-time PCR instruments. Master mix formulations optimized for different machines. 2x qPCR Master mix is ideally suited for:
•Gene expression analysis
•Microarray validation
•Viral load determination qRT-PCR
Kit Components
2x qPCR Master mix is a 2X mix of dNTPs, Hotstart Taq polymerase, MgCl2, fluorescent detection dye, reference dye(optional), and proprietary buffer components.
Storage
long time: -20°C
short time: 4°C
Recommended Protocol
Thaw 2x qPCR Master mix, template DNA, RNase-free water and primer on ice. Mix each solution well. Prior to the experiment, it is prudent to carefully optimize experiment Conditions and to include controls at every stage. See pre-protocol considerations for details. This standard protocol applies to a single reaction where only template, primers and water need to be added to the 2X qPCR Master mix. For multiple reactions, scale-up volume of reaction components proportionally. All reagents should be thawed on ice, gently mixed and briefly centrifuged before use.
1.Thaw reagents at room temperature. Mix thoroughly and then place on ice immediately after thawing.
2. Assemble reaction tubes on ice.
3. Prepare a reaction Master mix using the following:
A. Two-step fast cycling protocol
This cycling protocol should be applicable to most amplifications where the primer Tm’s are designed to be 60 oC. Melt curves may be performed by following instructions provided for your instrument.
This cycling protocol should be applicable to most amplifications where the primer Tm’s are designed to be 60 oC. Melt curves may be performed by following instructions provided for your instrument.
B. Three-step fast cycling protocol
This cycling protocol can be used if you would like to have the extension step to be performed at a higher temperature than the annealing step. For example, if you have relatively long primers that tend to anneal non-specifically, carrying out the extension step at a higher temperature can reduce nonspecific amplification. Melt curves may be performed by following instructions provided for your instrument.
This cycling protocol can be used if you would like to have the extension step to be performed at a higher temperature than the annealing step. For example, if you have relatively long primers that tend to anneal non-specifically, carrying out the extension step at a higher temperature can reduce nonspecific amplification. Melt curves may be performed by following instructions provided for your instrument.
Recommendations for Optimal Results
•Aliquot reagents to avoid contamination and to avoid repeated freeze-thaw cycles
•2x qPCR Master mix components are light sensitive; avoid exposure to light
•Start PCR as soon as the reaction mixture is prepared and always keep the reaction mixture chilled in an ice box prior to PCR reactions
NOTE:
1.Cycling conditions may need to be optimized, depending on different primer and template combinations. For example, raise the annealing temperature to prevent non-specific primer binding, increase extension time to generate longer PCR products.
2.Shorter annealing step time (<10sec) can be used for amplicon <100bp.
•Aliquot reagents to avoid contamination and to avoid repeated freeze-thaw cycles
•2x qPCR Master mix components are light sensitive; avoid exposure to light
•Start PCR as soon as the reaction mixture is prepared and always keep the reaction mixture chilled in an ice box prior to PCR reactions
NOTE:
1.Cycling conditions may need to be optimized, depending on different primer and template combinations. For example, raise the annealing temperature to prevent non-specific primer binding, increase extension time to generate longer PCR products.
2.Shorter annealing step time (<10sec) can be used for amplicon <100bp.
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