Product Number: PCM13B

Shipping and Storage

-20°C. Store at 2-8°C for frequent use.

Components

Note: The 2×Hotstar PCR Master Mix, with blue dye contains Golden Star Taq DNA Polymerase, 3.4mM MgCl₂ and 400µM

each dNTP.

Description

2×Hotstar PCR Master Mix is a premixed system consisting of Golden Star Taq DNA Polymerase, PCR Buffer, Mg²⁺, dNTPs, PCR stabilizers and reinforcers. The premixed PCR mixture makes the operation easier and faster. Minimizes human error and pollution. The Golden Star Taq DNA Polymerase is a chemically modified, new high efficiency Taq DNA Polymerase that completely blocks the enzyme activity at room temperature, making the enzyme inactive at low or normal temperatures. In order to effectively avoid the non-specific amplification caused by the non-specific combination of primer and template or primer dimer at room temperature, the activation of the enzyme must be incubated at 95°C for 10 min. The unique buffer system enables the enzyme to be widely used, enabling efficient amplification of templates with high GC content, complex secondary structure and low copy. The unique Master Mix formula makes the whole reaction system more stable. PCR amplification with this product, PCR product 3′ end with an “A” base, can be directly used for T/A cloning. This product does not contain dye, PCR procedure after bunching can be added according to the need of sample loading buffer after electrophoresis operation. This product has strong specificity and can be directly used for downstream cloning or chip hybridization experiments without the need of agarose gel recovery after PCR amplification. It is mainly used for conventional PCR, RT-PCR, multiple PCR and gene chip detection, especially for PCR reaction with high specificity requirements.

Quality Control

No exogenous nuclease activity was detected. No host residual DNA was detected by PCR. It can effectively amplify single copy genes in human genome. 2-8°Cstore three months, no significant activity change.

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.

Description

The Hot Start Taq DNA Polymerase is a recombinant Taq DNA polymerase which has been chemically modified by the addition of heat-labile blocking groups to its amino acid residues. The enzyme 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 need to be activated in incubation at 95°C for 10 minute. The activated enzyme maintains the same functionality.

Taq DNA polymerase: Has the 5′ to 3′ DNA polymerase activity and 5′ to 3′ exonuclease activity without 3′ – 5′ exonuclease activity. The extending speed is 1 kb/30 sec. There is an “A” on 3′ end. The PCR product can be cloned in TA vector.

Applications

• Hot-start PCR amplification
• Specific amplification of complex cDNA and genomic template
• Amplification from low copy number DNA template
• Real-Time PCR
• Multiple PCR
• Generation of PCR products for TA cloning, Gene microarray analysis and SNP test

Quality Control

• Functional absence of double and single-stranded endonuclease activity;
• Purity>99% test by SDS gel electrophoresis;
• Each lot of HotStar Taq DNA Polymerase is assayed for amplification from as little as 10 ng of human genomic DNA;
• Retain full activity at room temperature for one week;
• No host DNA residue.

Concentration: 5 u/μl

Package: Bulk

Storage: Store at -20°C

Order

Description

CSM Taq polymerases (Cold sensitive mutant Taq polymerases ) is a kind of HotStar Polymerase. Source from the mutant E. coli. CSM Taq polymerase unlike the monoclonal antibody based hot start Taq or chemical modified hot start Taq, Cold Taq is the cold sensitive mutant Taq. It retain the hot start ability throughout the whole amplification cycles.

Cold sensitive mutant Taq polymerases are designed for Hot Start PCR, it is not at low temperature. It offers excellent specificity and two-fold higher fidelity than wild-type Taq. It is designed for PCR with difficult templates such as GC-rich fragments and microsatellites.

Cold sensitive mutant Taq polymerases are particularly well suited to primer extension of Single Nucleotide Polymorphism (SNP) markers.

Cold sensitive mutant Taq polymerases maintain excellent specificity and minimal background even in conditions designed for high yield. In fact, even on genomic templates, the enzyme can be used with MgCl2+concentrations as high as 10 mM.

Cold sensitive mutant Taq polymerases are capable of extending through difficult regions, e. g. regions, which include inverted tandem repeats and those with high amounts of secondary structure.

Cold sensitive mutant Taq polymerases work in a totally unique way, involving improved nucleotide selection at the active site, and a much lower rate of mis-match extension, meaning that only perfectly aligned primers will be extended. As a result, the enzyme can give even higher specificity than hot-start (manual or automatic) techniques without the need for inconvenient pre-incubation steps.

Applications

• Hot start PCR amplification
• Specific amplification of complex cDNA and genomic template, for amplification of difficult templates, such as GC-rich fragments and microsatellites
• Primer extension of SNP markers
• Amplification of genomic DNA targets up to 10 kb with high fidelity, specificity, and sensitivity
• Amplification from low copy number DNA template, high through-put Hot Start PCR with high specificity, sensitivity, and yield
• Routine diagnostic Hot Start PCR requiring high reproducibility.
• Real-Time PCR
• Multiple PCR
• Generation of PCR products for TA cloning

Quality Control

• Functional absence of double and single-stranded endonuclease activity;
• Purity>99% test by SDS gel electrophoresis;
• Each lot of CSM Taq DNA Polymerase is assayed for amplification from as little as 10 ng of human genomic DNA ;
• Retain full activity at room temperature for one week;
• No host DNA residue.

Concentration: 5 U/μl

Storage: Store at -20°C

Package: Bulk

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.

The kit already include MMLV, HotStar DNA polymerase, RNAse inhibitor, dNTPs, MgCL and so on. Which special optimized for COVID-19 Virus. Customer only need add oligos, probe, and positive control and negative control then can be your own detection kit. This kit offered by bulk pack, easy for your make the detection kit. If need oligo, probe, template we also can offer.

Hot Star Probe One Step RT-qPCR Kit is designed for one-step qPCR based on probes (TaqMan, Molecular Beacon etc.). The reverse transcription and quantitative PCR are carried out in the one reaction system, no need to add other reagents in the whole reaction process. The unique reverse transcriptase in the kit greatly improved transcriptional activity. The new hot start polymerase has the advantages of high amplification efficiency and good specificity. The special buffer system contained in the kit can make the reverse transcriptase and DNA polymerase play the maximum role at the same time and improve the reaction efficiency. GoldStar Probe One Step RT-qPCR Kit is highly sensitive, strong fluorescent signal and highly signal-to-noise ratio. It is very suitable for detecting RNA such as RNA virus, new coronavirus (20I9-nCoV). Using this product, we can get a wider linear dynamic range, more accurate quantitative target gene, good repeatability and high reliability.

Feature:

• (1) One Step RT-PCR allows accurate, rapid analysis of small amounts of RNA such as viral RNA.
• (2) Hotstar Polymerase enables highly efficient, specific amplification and detection.
• (3) One Step RT-PCR Buffer is a 2X premix, which allows simple preparation of the reaction with minimal risk of contamination.
• (4) easy to use, the kit already include MMLV, HotStar DNA polymerase, RNAse inhibitor, dNTPs, MgCL and so on. Special optimized for COVID-19. Customer only need add oligos, probe, and positive control and negative control then can be your own detection kit. If need oligo,probe, template we also can offer.

Quality Control:

Function test by RT-qPCR use WHO and CDC Recommended template, and primer

Product Number: PCM13

Shipping and Storage

-20°C. Store at 2-8°C for frequent use.

Components

Description

2×Hotstar PCR MasterMix is a premixed system consisting of Golden Star Taq DNA Polymerase, PCR Buffer, Mg²⁺, dNTPs, PCR stabilizers and reinforcers. The premixed PCR mixture makes the operation easier and faster. Minimizes human error and pollution. The Golden Star Taq DNA Polymerase is a chemically modified, new highefficiency Taq DNA Polymerase that completely blocks the enzyme activity at room temperature, making the enzyme inactive at low or normal temperatures. In order to effectively avoid the non-specific amplification caused by the non-specific combination of primer and template or primer dimer at room temperature, the activation of the enzyme must be incubated at 95°C for 10 min. The unique buffer system enables the enzyme to be widely used, enabling efficient amplification of templates with high GC content, complex secondary structure and low copy. The unique MasterMix formula makes the whole reaction system more stable. PCR amplification with this product, PCR product 3′ end with an “A” base, can be directly used for T/A cloning. This product does not contain dye, PCR procedure after bunching can be added according to the need of sample loading buffer after electrophoresis operation. This product has strong specificity and can be directly used for downstream cloning or chip hybridization experiments without the need of agarose gel recovery after PCR amplification. It is mainly used for conventional PCR, RT-PCR, multiple PCR and gene chip detection, especially for PCR reaction with high specificity requirements.

Quality Control

No exogenous nuclease activity was detected. No host residual DNA was detected by PCR. It can effectively amplify single copy genes in human genome. 2-8°Cstore three months , no significant activity change.