2x Ultra SYBR Mixture (With separate ROX) is a convenient premixed 2x concentrated solution for SYBR Green I-based qPCR which include HotStar Taq DNA polymerase, PCR buffer, dNTPs, SYBR Green I fluorescent dyes, Mg²⁺. It can be applied to the target sequence detection of genomic DNA and cDNA. The SYBR Green I dye binds to dsDNA without using sequence-specific probes. The modified HotStar Taq DNA polymerase is inactive under 40°C which effectively avoids the non-specific amplification caused by primer dimers or the nonspecific binding of prime and template. The unique PCR buffer components and the hot start enzyme ensure PCR specificity and sensitivity. The ROX is a dye molecule that can be used to normalize the well-to well fluorescent fluctuations and can be applied to all qPCR instruments using ROX as calibration dye. It has a higher sensitivity and a wider linear range which can exactly quantitate the target gene or low copy template.

Packing

2x SYBR qPCR mix 1.25ml or bulk
Rox reference dye (50µmol) 25ul

Application

Gene expression analysis
Gene copy number analysis

Features

High sensitivity: exactly quantitate the low copy template
High specificity: maximatily reduce the primer dimers and non-specific product
Wide linear range; Accurate quantitation

Shipping And Storage

Store in dark at -20°C for up to 1 year.
For frequent use, Store at 2-8°C for short-term (1 week), and avoid freeze thawing.

Rox reference dye user guide

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Storage

Store at -20 ℃ and transport on dry ice.

Components

Description

The 2×Ultra Sybr qPCR Mix (Low Rox) is a premixed system for realtime fluorescence quantitative PCR (SYBR Green I), and the concentration is 2×. It contains Golden Star Taq DNA Polymerase, PCR Buffer, dNTPs, SYBR Green I Fluorescent Dye, Mg²⁺ and Low ROX as reference dye. The operation is simple and convenient. This product is mainly used for the detection of genomic DNA target sequences and cDNA target sequences after RNA reverse transcription.

This product contains the fluorescent dye SYBR Green I which can bind with all double-strand DNA, so that the product can be used for the detection of different target sequences without the need for the synthesis of specific labeled probes. The Golden Star Taq DNA Polymerase in the mixture is a chemically-modified, new efficient hot-start enzyme that does not have polymerase activity at room temperature which prevents non-specific amplification efficiently, and it is activated by incubation at 95°C for 10 minutes. The combination of a unique PCR buffer system and a hot-start enzyme effectively inhibits non-specific PCR amplification and significantly increases the amplification efficiency of PCR.

The ROX dye contained can correct the fluorescence signal error between the wells of the quantitative PCR instrument. The amount of ROX in this kit is low, and it is suitable for quantitative PCR instruments which require low ROX for signal correction, such as ABI Prism 7500/7500 Fast, Stratagene Mx3000/MX3005P, and Corbett Rotor Gene 3000.

Features

1. This product uses a new high-performance hot start enzyme (Golden Star Taq DNA Polymerase) and a unique PCR buffer system. This product significantly improves the PCR amplification efficiency and has high sensitivity and specificity.
2. This product is suitable for quantitative PCR detection and can accurately quantify and detect the target gene.

Application

• Gene expression analysis
• Gene copy number analysis

Order

The 2x TaqMan qPCR master (low rox) Mix is a special mixture for Probe real time PCR(TaqMan , Molecular Beacon etc.). Which include the HotStar DNA Polymerase, PCR Buffer, MgCl2 , dNTPs, ROX and the stabilizing agent. The HotStarTaq DNA polymerase which in the mix is inactive at room temperature, avoiding extension of non-specifically annealed primers or primer dimers and providing higher specificity of DNA amplification. The functional activity of the enzyme is activated in 10 minute incubation at 95°C. The unique reaction buffer with the special Hotstar enzyme highly increased the Amplification efficiency. The fluorescence signals is stronger, the sensitivity is higher can detect the Single copy sequences. The Rox in the mix can be used for the correction of the fluorescent signal in the qPCR. The 2x TaqMan qPCR master Mix produces a optimal results in qPCR.

The kit is applied to a wide linear range and can exactly quantitate the target gene. It can be widely applied to all qPCR instruments without using ROX as calibration dye. The applicable machine as the flowing: ABI Prism 7500/7500 Fast, QuantStudio® 3 System, QuantStudio® 5 System, QuantStudio® 6 Flex System, QuantStudio® 7 Flex System, ViiA 7 system, Stratagene Mx3000/Mx3005P, Corbett Rotor Gene 3000

Note:

1.Before use, please upside down gently for blending, avoid foam, and short centrifuge.
2.The Master Mix contain the ROX need to avoid the hard light when use and storage.
3.Avoid repeated freezing and thawing, frequently use can be stored at2-8°C, for long term storage can be store at -20°C.

Protocol:

Reaction Mixture Set Up

Note:

1.Generlly the primer conc at 0.2 μM can get a good result, the user can try from 0.1μ-1.0μM.
2.The conc of the Probe should be adjust according the actuality. User can refer to the instruction of instrument and the probe.
3.Usually user can refer to 10-100ng genomic DNA or 1-10ng cDNA as the template, but as different template have different copies, user should make a Gradient dilution to confirm the optimum conc.
Recommended thermal cycling conditions
95 °C 10 min
95 °C 15 sec 30-45 cycles
60 °C 60 sec 30-45 cycles

Note:

1.The functional activity of the enzyme should to be activated in 10 minute incubation at 95°C
2.We recommend use the two step PCR, but if can not get the good result caused by the low Tm primer and other reasons, user can use the three steps PCR, and the anneal temperature we suggest 56-60°C

Related Products:

We also have oligo, hot star taq, proteinase K and PCR and so on! If you need more , Please contact us as you need.

Product Number: PCM40, PCM41

Shipping and Storage

-20°C; If used frequently, store at 2-8°C, avoiding repeated freezing and thawing.

Components

Description

The 2×Taq Man qPCR MasterMix is a premixed system for qPCR based on probes (TaqMan, Molecular Beacon etc.), and the concentration is 2×. It contains GoldenStar Taq DNA Polymerase, PCR Buffer, dNTPs, and Mg²⁺. The operation is simple and convenient. This product is mainly used for the detection of genomic DNA target sequences and cDNA target sequences after RNA reverse transcription, such as gene expression analysis, copy number analysis, SNP genotyping, etc., and is applicable to the qPCR using different types of probes.

The Golden Star Taq DNA Polymerase in the mixture is a chemically-modified, new efficient hotstart enzyme that does not have polymerase activity at room temperature which prevents nonspecific amplification efficiently, and it is activated by incubation at 95°C for 10 minutes. The combination of a unique PCR buffer system and a hot-start enzyme significantly increases the amplification efficiency of PCR. The fluorescence signal is stronger, and it is more sensitive, which can even detect single copy template. This product can be used to get a wider linear range and more accurate quantification of the target gene. This product is suitable for fluorescent qPCR instruments that do not require ROX as a calibration dye.

Notes

1. Mix gently before use, avoid foaming, and use after brief centrifugation.
2. Avoid repeated freezing and thawing of this product. Repeated freezing and thawing may comprise product performance. This product can be stored at -20°C in dark for long-term storage. If used frequently in a short time, it can be stored at 2-8°C.