2× Medino® HotStart PCR Master Mix

Product2× Medino® HotStart PCR Master Mix
Catalog No.LM201007-3
Storage conditionsThis product should be stored at -20℃ for 2 years.

Description

Product description

2× Medino® HotStart PCR Master Mix is a ready-to-use PCR premixed solution containing Medino® HotStart Taq DNA Polymerase, dNTP and optimized buffer system, which can be amplified only by adding primers and templates, which greatly simplifies the operation steps of the experiment, can be high-throughput operation and improve the reproducibility of the experimental results. The activation time of Medino® HotStart DNA Polymerase takes only 2-3 min, which is compatible with existing PCR programs. This product prevents non-specific amplification during the sample preparation stage and can effectively perform gene cloning and typing experiments. The PCR product with 3 ′-dA, protruding end, can be easily cloned into the T vector. The premix solution contains no dye.

Instructions

  1. Recommended PCR reaction systems.
ComponentsVolume (μL)Final concentration
ddH2OUp to 50
2× Medino® HotStart PCR Master Mix25
Templatex
Forward Primer 10 μmol/L20.4 μM
Reverse Primer 10 μmol/L20.4 μM
[Note]:For the different optimal reaction concentration of different templates, the following table is the recommended template usage of 50 μL reaction system, This is for your reference only. Recommended usage of the different templates:
Template typesTemplate usage
Genome DNA50 ng – 200 ng
Plasmid DNA100 pg – 20 ng
cDNA1-5 μL (no more than the reaction system 1/10)
  1. Reaction program
Cycle stepTemp.TimeCycles
Initial denaturation95℃5 min1
Denaturation95℃30 sec35
Annealing50-60℃30 sec
Extension72℃30 sec/kb
Final extension72℃10 min1
[Note]:
  1. Annealing temperature and time: 50 – 60℃ is recommended for the temperature. Too low annealing temperature causes non-specific amplification. Primers were designed with reference to fluorescence quantitative primer standards. The recommended annealing time is set to 30 sec, which can be adjusted within 10 – 30 sec. Too long an annealing time may cause the dispersion of the amplification product on the glue. Can also Set up a temperature gradient to explore the optimal temperature and time of primer annealing as needed.
  2. Extended temperature and time: 72℃ of temperature is recommended.30 sec/kb is recommended.
  3. Amplification product: Please place the PCR amplification product at -20℃ for storage to prevent DNA degradation.

Notes

  1. a) For your safety and health, please wear a lab coat and disposable gloves during
  2. b) This product is for research use only!

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