2 × Medino® HotStart PCR Genotyping Master Mix (With Dye)

 

Cat.No. LM201007-2
Size1 mL/5×1 mL/50×1 mL/100×1 mL
Storage and shippingThe product is shipped with dryice and can be stored at -20℃ for  2 year.
ApplicationFor mouse genotyping mainly
QCExonuclease 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.

Category:

PDF download: LM201007-2

 

Product description

2 × Medino® HotStart PCR Genotyping Master Mix (With Dye) is a ready-to-use PCR premix solution, containing Medino® 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. Medino® 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. Medino® 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 μLFinal concentration
2 × Medino® HotStart PCR Genotyping Master Mix (With Dye)251 ×
Templatex
Forward Primer (10 μmol/L)20.4 μmol/L
Reverse Primer (10 μmol/L)20.4 μmol/L
ddH20Up 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 DNA50 ng-100 ng
Plasmid DNA100 pg-20 ng
cDNA1-5 μL (no more than 1/10 of the reaction system)

2.Reaction program

Cycle stepTemp.TimeCycles
Initial denaturation95 °C5 min1
Denaturation95 °C30 sec35
Annealing50-60 °C30 sec
Extension72 °C30 sec
Final extension72 °C10 min1

[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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