2× Medino® Ultra-Long PCR Master Mix-with Dye

Product2× Medino® Ultra-Long PCR Master Mix-with Dye
Catalog No.LM201008-2
Storage conditionsThis product should be stored at -25~15℃ for 1years.

Description

Product description

2× Medino® Ultra-Long PCR Master Mix (With Dye) has ligand-modified heat-stabilized Taq DNA Polymerase, fused 3 ′-5 ′-corrected activity factor and optimized buffer system, able to amplify 25 kb of genome fragments, 14 kb of cDNA gene sequence, and a 40 kb λ DNA fragment. The dNTP and Mg2+ required for amplification are included in Mix. When used, only primers and template can be used, avoiding various losses caused by the addition process. Meanwhile, Mix also contains the dye part, which greatly reduces the tedious steps of subsequent electrophoresis. In addition, the protective agent added to the system enables the product to be maintained after repeated freezing and thawing Stable activity. PCR, the 3 ′ end of the product, band A, can be directly ligated to the T vector.

Instructions

  1. Recommended PCR reaction systems.
ComponentsVolume (μL)Final concentration
ddH2OUp to 50
2× Medino® Ultra-Long PCR Master Mix-with Dye25
Templatex
Forward Primer 10 μmol/L20.4 μM
Reverse Primer 10 μmol/L20.4 μM
[Note]:Reaction buffer: The buffer contains 2.5 mM Mg2+ and the optimized dNTPs components. At the same time, the kit provides additional MgSO4 and dNTPs. For fragments with difficult amplification above 20 kb, we can try to add Mg2 + and 200-500 μM. 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℃3 min1
Denaturation98℃10 sec35
Annealing68℃30 sec/kb
Final extension72℃10 min1
[Note]:
  1. Annealing / extension: Expansion of the growing fragments is recommended using a two-step method. The annealing / extension time was set at 30 sec/kb to amplify most of the fragments, and to prolong the reaction appropriately for fragments above 20 kb Time, should not exceed 60 sec/kb

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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