Product Number: PCM47

Storage Condition

-20±5℃.

Components

Description

SuperStar sProbe Mixture is a premix solution dedicated to real-time fluorescent quantitative PCR using probe methods (TaqMan, Molecular Beacon, etc.). The core component SuperStar DNA Polymerase is a hot-start DNA polymerase modified by double antibodies. It can recover DNA polymerase activity by heating at 95°C for 5 seconds. It has many advantages such as strong specificity and high detection sensitivity, and is equipped with the most suitable buffer optimized for qPCR. , the concentration of the reaction solution was 2×. The unique qPCR buffer system and hot-start enzyme combination of this product can effectively inhibit the generation of non-specific products and significantly improve the amplification efficiency of qPCR. It is very suitable for high specificity, high sensitivity, single and multiplex amplification, etc. qPCR reaction. Customers only need to add templates, primers, and probes, which is easy to use.

Pre-experiment preparation and precautions

• 1. Before use, please mix the product upside down and gently after it is completely melted, and use it after a short centrifugation.
• 2. This product is recommended to be used in small portions. If it needs to be used frequently in a short period of time, it can be stored at 2-8°C for 6 months.
• 3. Avoid repeated freezing and thawing of this product. Repeated freezing and thawing (freezing and thawing times ≤ 20 times) may reduce the performance of the product.
• 4. This product can be stored at -20±5°C and protected from light for a long time.
• 5. When configuring the reaction system of this product, it is best to carry out in a clean bench or a sterile environment.

Product Number: PCM63, PCM63L, PCM63H

Storage Condition

-20°C. For frequent use, it can be stored at 2-8°C. Try to avoid repeated freezing and thawing.

Components

Description

2×HotStar Probe Mix UNG is a premix system dedicated to real-time fluorescent quantitative PCR with a probe method (TaqMan, Molecular Beacon, etc.), with a concentration of 2×, including HotStar Taq DNA polymerase, PCR Buffer, dNTPs (dTTP is all replaced by dUTP replaced), UNG enzyme and Mg2+, the operation is simple and convenient. It 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 genotype analysis, etc. The dUTP-UNG anti-pollution system is used in this product, and dUTP is added in the preparation process of the PCR reaction system, so an amplification product containing dU bases is formed. This product can be eliminated by UNG enzyme treatment in the PCR system before the next PCR reaction. In this way, the residual contamination of the PCR product is effectively removed, and the false positive caused by the contamination of the amplification product is greatly reduced. The UNG enzyme can be inactivated by the pre-denaturation step in the PCR cycle, so it will not affect the formation of new dU-containing PCR products. The HotStar Taq DNA Polymerase contained in this product is a chemically modified, brand-new high-efficiency hot-start enzyme, which has no polymerase activity at room temperature, and effectively avoids non-specific binding of primers and templates or primer dimers at room temperature. To generate non-specific amplification, enzyme activation must be incubated at 95°C for 10 minutes. The unique combination of PCR buffer system and hot-start enzyme significantly improves the amplification efficiency of PCR, with stronger fluorescent signal and higher sensitivity, and can detect single-copy templates. Using this product can get a wider linear range and more accurate quantification of the target gene.

ROX dye is used to correct the fluorescence signal error between the wells of the quantitative PCR instrument, and is generally used in Real Time PCR amplification instruments of companies such as ABI and Stratagene. The excitation optics of different instruments are different, so the concentration of ROX dye must be matched with the corresponding real-time PCR instrument.

Instruments that do not require ROX calibration (PCM63):

Roche LightCycler 480, Roche LightCyler 96, Bio-radi iCyler iQ, iQ5, CFX96, etc.

Instruments requiring Low ROX calibration (PCM63L):

ABI Prism7500/7500 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 etc.

Instruments requiring High ROX calibration (PCM63H):

ABI Prism7000/7300/7700/7900, Eppendorf, ABI Step One/Step One Plus etc.

Note

1. Before use, please mix it upside down gently, try to avoid foaming, and use it after short centrifugation.

2. Avoid repeated freezing and thawing of this product, as repeated freezing and thawing may reduce product performance. Long-term storage of this product can be stored at -20°C, away from light. If it needs to be used frequently in a short period of time, it can be stored at 2-8°C.

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: PCM39

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.

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.