Conventional PCR

Product description The kit is a ready-to-use PCR premixed solution containing HotStart Taq DNA Polymerase, dNTPs, and an optimized buffering system. Simply add primers and template for amplification, which greatly simplifies the experimental procedure and allows for high-throughput operations, enhancing the reproducibility of results. The hotStart Taq DNA Polymerase is a ligand-modified thermostable Taq DNA Polymerase, with a ligand that modulates the activity of the DNA polymerase in response to temperature changes. The enzyme activity is completely sealed at room temperature and is only released after heating at 95℃. The activation time for HotStart DNA Polymerase requires only 2-3 min and is compatible with existing PCR protocols. This product prevents non-specific amplification during sample preparation and reaction heating stages, effectively enabling genotyping experiments. Components Name N132004E N132004S N132004M N132004L 2×HotStart Genotyping PCR Master Mix (With Dye) 1 mL 5×1 mL 50×1 mL 100×1 mL Specifications Concentration 2× Hot Start Built-in Hot Start Overhang 3 '-A Polymerase Taq DNA Polymerase Reaction speed Standard Product Type PCR Master Mix (2x) Shipping and Storage Dry ice shipping. -15℃ ~ -25℃ storage, valid for two years. Applications This product is primarily applied to the genotyping of mice. Notes 1. For your safety and health, please wear lab coats and disposable gloves for operation. 2. This product is for research use ONLY!This product is for research use ONLY! Instructions 1. Reaction System  Table 1 Reaction system Components Size (μL) Final concentration 2×HotStart Genotyping PCR Master Mix (With Dye) 25 1× Template DNA suitable - Forward primer (10 μmol/L) 2 0.4 μM Reverse primer (10 μmol/L） 2 0.4 μM ddH2O to 50 - Recommended usage of the different templates: Table 2 Recommended usage of different templates Type of template Segment usage range (50 μL reaction system) Genomic DNA 50–100 ng Plasmid DNA 0.1-20 ng cDNA 1-5 μL (Do not exceed 1/10 of the reaction system)   2. Amplification Protocol Table 3 Amplification protocol Cycle steps Temperature (°C) Time Cycles Predenaturation 95 5 min 1 Denaturation 95 30 sec 35 Annealing 50-60 30 sec Extension 72 30-60 sec/kb Final extension 72 10 min 1 【Note】: a. *Recommended Annealing Temperature and Time: The annealing temperature is recommended to be set at 50-60℃. The annealing time is recommended to be set at 30 sec, with adjustments possible within the range of 20-30 sec. Depending on the need, a temperature gradient can be established to explore the optimal annealing temperature and time for primer annealing. b. **Extension Temperature and Time: The temperature is recommended to be 72℃. The time is recommended to be 30-60 sec/kb. c. **PCR Amplification Products: Please store the PCR amplification products at -20℃ to prevent DNA degradation.

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Product description The kit is a kind of conventional PCR premixed solution which is ready to use, including Taq DNA Polymerase, dNTP mix, MgCl2 and optimized buffer. During the reaction, only the primer and template can be added for amplification, which greatly simplifies the operation steps of experiment. This product contains excellent stabilizers and can be stored for 3 months at 4℃. The PCR product have 3 '-dA protrusion and can be easily cloned into T vector. Components Components No. N132003E N132003S N132003M N132003L Size 1 mL 5×1 mL 50×1 mL 100×1 mL Specifications Fidelity (vs. Taq) 1× Hot Start No Overhang 3 '-A Polymerase Taq DNA Polymerase Reaction Format SuperMix or Master Mix Reaction Speed Standard Product Type PCR Master Mix (2x) Shipping and Storage Dry ice shipping. -15℃ ~ -25℃ storage, valid for two years. Notes 1. PCR products with 2×PCR Master Mix are not suitable for polyacrylamide gel electrophoresis. 2. For your safety and health, please wear lab coats and disposable gloves for operation. 3. This product is for research use ONLY! Instructions 1. Reaction System  Table 1  Reaction system Components Size (μL) Template DNA suitable Primer 1 (10 μmol/L) 2 Primer 2 (10 μmol/L） 2 2×PCR Master Mix 25 ddH2O to 50 【Note】: 1) Template usage: 50-200 ng genomic DNA; 0.1-10 ng plasmid DNA. 2) Mg2+ concentration: This product contains 3 mM of MgCl2, suitable for most PCR reactions. 2. Amplification Protocol Table 2  Amplification protocol Cycle steps Temperature (°C) Time Cycles Predenaturation 94 5 min 1 Denaturation 94 30 sec 35 Annealing 50-60 30 sec Extension 72 30-60 sec/kb Final extension 72 10 min 1 【Note】: a. Annealing temperature: Please refer to the theoretical Tm value of primers. The annealing temperature can be set to 2-5℃ lower than the theoretical value of the primer. b. Extention time: For molecular identification, 30 sec/kb is recommended. For gene cloning, 60 sec/kb is recommended. Application example   Figure 1 The expected 1.2 kb PCR products can be amplified with 2× PCR Master Mix. The Master Mix was stored at -20℃ for 1 year following another 3 months at 4℃ and 1 month at 25℃. Template: Arabidopsis genome. Annealing temperature: 60℃. Extension time: 40 sec.  

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Product description The kit contains Taq DNA Polymerase, dNTPs, and other PCR-required components. The Master Mix is stable for 3 months at 4℃ with our customized stabilizers. The pre-mix solution is optimized for conventional PCR and ready to use by adding DNA template and primers. The PCR products can be loaded directly for electrophoresis with pre-loaded bromophenol blue dye. The amplified products contain 3 '-dA protrusion and can be easily cloned into T vector. The 2×PCR Master Mix simplifies PCR procedure and reduces contamination. Components Components No. N132002E N132002S N132002M N132002L Size 1 mL 5×1 mL 50×1 mL 100×1 mL Specifications Fidelity (vs. Taq) 1× Hot Start No Overhang 3 '-A Polymerase Taq DNA Polymerase Reaction Format SuperMix or Master Mix Reaction Speed Standard Product Type PCR Master Mix (2x) Shipping and Storage Dry ice shipping. -15℃ ~ -25℃ storage, valid for two years. Notes 1. PCR products with 2×PCR Master Mix are not suitable for polyacrylamide gel electrophoresis. 2. For your safety and health, please wear lab coats and disposable gloves for operation. 3. This product is for research use ONLY! Instructions 1. Reaction System   Table 1  Reaction system Components Size (μL) Template DNA suitable Primer 1 (10 μmol/L) 2 Primer 2 (10 μmol/L） 2 2×PCR Master Mix 25 ddH2O to 50 【Note】: 1) Template usage: 50-200 ng genomic DNA; 0.1-10 ng plasmid DNA. 2) Mg2+ concentration: This product contains 3 mM of MgCl2, suitable for most PCR reactions. 2. Amplification Protocol Table 2  Amplification protocol Cycle steps Temperature (°C) Time Cycles Predenaturation 94 5 min 1 Denaturation 94 30 sec 35 Annealing 50-60 30 sec Extension 72 30-60 sec/kb Final extension 72 10 min 1 【Note】: a. Annealing temperature: Please refer to the theoretical Tm value of primers. The annealing temperature can be set to 2-5℃ lower than the theoretical value of the primer. b. Extention time: For molecular identification, 30 sec/kb is recommended. For gene cloning, 60 sec/kb is recommended. Application example   Figure 1 The expected 1.2 kb PCR products can be amplified with 2× PCR Master Mix. The Master Mix was stored at -20℃ for 1 year following another 3 months at 4℃ and 1 month at 25℃. Template: Arabidopsis genome. Annealing temperature: 60℃. Extension time: 40 sec.  

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Product description 2× HotStart PCR Master Mix contains heat-stabilized Taq DNA Polymerase modified with antibodies, adds strong elongation factor and optimized buffer system, and has super high amplification efficiency. It is very suitable for PCR amplification of most colonies such as E. coli, Agrobacterium, and yeast. The extension speed of amplification of complex templates within 3 kb can reach 1 sec/kb, 3-6 kb to 3 sec/kb, 6-10 kb to 5 sec/kb and 10 kb to 10 sec/kb; the amplification speed of simple templates such as plasmid within 6 kb can reach 1 sec/kb, which can greatly save PCR reaction time. Meanwhile, Mix contains dNTPs, Mg2+, and it can be amplified only with adding primers and template. In addition, Mix contains red tracer dye, which can be used by electrophoresis directly after the end of the reaction. The protective agent added to the system enables the product to maintain the stable activity after repeated freezing and thawing. The 3′   end of the PCR product bands A and can be easily cloned into the T vector. Components Components No. N132001E N132001S N132001M N132001L Size 1 mL 5×1 mL 50×1 mL 100×1 mL Specifications Product specification Master Mix Concentration 2× Hot Start Built-in Hot Start Overhang 3 '-A Reaction speed Rapid Size (Final Product) Up to 15 kb Conditions for transportation Dry ice Shipping and Storage Dry ice shipping. -15℃ ~ -25℃ storage, valid for two years. Notes 1.  For your safety and health, please wear lab coats and disposable gloves for operation. 2. This product is for research use ONLY! Instructions 1. Reaction System  Table 1  Reaction system Components Size (μL) Size (μL) Final concentration 2×HotStart Fast PCR Master Mix (With Dye)* 25 12.5 1× Template DNA** suitable suitable - Forward primer (10 μmol/L) 2 1 0.4-0.5 μM Reverse primer (10 μmol/L） 2 1 0.4-0.5 μM ddH2O to 50 to 25 - 【Note】: 1) *The 2×Mix contains 2 mM Mg2+ and 200 μM dNTPs, thawed thoroughly before use. 2) **E. coli and Agrobacterium can directly absorb bacterial liquid or pick bacteria samples; It is recommended that the yeast liquid and 3) **Strain be boiled for 5 min, then put into the -80℃ refrigerator for 3 min, and then serve the sample after thawing. Note that the bacterial liquid sample should be shaken and mixed before sampling, in which the recommended amount of bacterial liquid sample is 2-4 μL (0.5-0.8 OD600). Recommended usage of the different templates: Table 2  Recommended usage of different templates Type of template Segment usage range (25 μL reaction system) Genomic DNA 10–1,000 ng Plasmid or  λDNA 0.5-50 ng E. coli bacteria solution 0.5-0.8 OD600 【Note】: ***The range of final primer concentration in PCR reaction system is 0.2-1 μM, and 0.4 μM is recommended. 2. Amplification Protocol Table 3  Amplification protocol Cycle steps Temperature (°C) Time Cycles Predenaturation 95 3 min 1 Denaturation 95 15 sec   30-35 Annealing* 60 20 sec Extension** 72 1-10 sec/kb Final extension 72 5 min 1 【Note】: a. *Recommended annealing temperature: 60°C, you can also set up a temperature gradient according to your own needs to explore the optimal temperature for primer annealing. The recommended annealing time is set to 20 sec, which can be adjusted within 10-30 sec. Too long annealing time may lead to dispersion of the amplified product on the glue. b. **Extension speed: 1sec/kbfor complex templates such as genomes and E. coli within 3 kb, 3 sec/kb for complex templates within 6 kb, 5  sec/kb for most complex templates within 10 kb, and 10 sec/kb for complex template fragments over 10 kb. For simple templates, such as plasmids less than 6 kb, set 1 sec/kb; for simple templates, such as 6-10 kb plasmids, set 3 sec/kb; and for simple templates, such as plasmids larger than 10 kb, set 5-10 sec/kb. If it is necessary to increase production, the extension time can be extended appropriately, and should not exceed 30 sec/kb.  

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