詳細(xì)說(shuō)明
Lolina A/SAddress: Sindalsvej 30 8240 Risskov Danmark
Email: Info@lolina.dk
Website: https://lolina.dk
Product Specification
Product name | 2 × Lolina? HotStart PCR Genotyping Master Mix (With Dye) |
Cat.No. | NaM201007-2 |
Size | 1 mL/5×1 mL/50×1 mL/100×1 mL |
Storage and shipping | The product is shipped with dryice and can be stored at -20℃ for 2 year. |
Application | For mouse genotyping mainly |
QC | Exonuclease residue detection: 20 μL of this product and 0.6 μg λ DNA-Hind III were incubated for 4 h at 37°C. There was no change in the electrophoresis band of DNA. Endonuclease residue detection: 20 μL of this product and 1 μg of λDNA were incubated at 37°C for 4 hours. There was no change in the DNA electrophoresis band. Detection of Escherichia coli residual DNA: Add 25 μL of this product to a 50 μL system, and use sterile ddH2O as a template to amplify the E.coil 16s rDNA gene. After 30 cycles, the amplified products were subjected to 1% agarose gel electrophoresis and EB staining, and no amplified bands were found. |
Product description
2 × Lolina? HotStart PCR Genotyping Master Mix (With Dye) is a ready-to-use PCR premix solution, containing Lolina? HotStart Taq DNA Polymerase, dNTPs and an optimized buffer system. Just add primers and templates for amplification, greatly simplifying the experimental steps and enabling high-throughput operation and improve the reproducibility of experimental results. Lolina? HotStart Taq DNA Polymerase is a thermostable Taq DNA Polymerase modified with a ligand that modulates DNA polymerase activity as a function of temperature. Enzyme activity is completely blocked at room temperature and is released after heating to 95°C. Lolina? HotStart DNA Polymerase requires only 2-3 minutes to activate and is compatible with existing PCR protocols. This
product prevents non-specific amplification during sample preparation and reaction heating stages, and can effectively perform genotyping experiments.
Instructions
1. Recommended PCR Reaction System (50 μL)
Components | Volume μL | Final concentration |
2 × Lolina? HotStart PCR Genotyping Master Mix (With Dye) | 25 | 1 × |
Template | x | - |
Forward Primer (10 μmol/L) | 2 | 0.4 μmol/L |
Reverse Primer (10 μmol/L) | 2 | 0.4 μmol/L |
ddH20 | Up to 50 | - |
[Note]:
The optimal reaction concentration for different templates is different. The following table is the recommended template usage for a 50 μL reaction system, which is for reference only.
Components | Volume μL |
Genomic DNA | 50 ng-100 ng |
Plasmid DNA | 100 pg-20 ng |
cDNA | 1-5 μL (no more than 1/10 of the reaction system) |
2. Reaction program
Cycle step | Temp. | Time | Cycles |
Initial denaturation | 95 °C | 5 min | 1 |
Denaturation | 95 °C | 30 sec |
35 |
Annealing | 50-60 °C | 30 sec | |
Extension | 72 °C | 30 sec | |
Final extension | 72 °C | 10 min | 1 |
[Note]:
1) Annealing temperature and time: The recommended annealing temperature is 50-60°C. The recommended annealing time is 30 sec, which can be adjusted within 20-30 sec. As needed, a temperature gradient can be set up to find the optimal temperature and time for index annealing.
2) Extension temperature and time: The recommended temperature is 72°C. The recommended time is 30-60 sec/kb.
3) Amplification product: Please store the PCR amplification product at -20°C to prevent DNA degradation.
Notes
1. This product is for research use only.
2. Please operate with lab coats and disposable gloves for your safety.
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