NaM201007-2_2 × Lolina HotStart PCR Genotyping Master Mix (With Dye)
详细说明
Lolina A/S
Address: 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.
产品详细信息请联系产品工程师
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