2x Medino® Ultra-Long PCR Master Mix

 

Catalog No.LM201008-1
Storage conditionsDry ice transportation. Store at -20°C with a shelf life of 1 year.

Product description

2× Medino® Ultra-Long PCR Master Mix contains ligand-modified thermostable Taq DNA Polymerase fused with a 3’-5’ exonuclease activity factor, as well as optimized buffer system. It can amplify genome fragments up to 25 kb, cDNA gene sequences up to 14 kb, and λDNA fragments up to 40 kb. The mix includes dNTPs and Mg2+ required for amplification, eliminating various losses during the addition process. Additionally, protective agents in the system maintain stable activity even after repeated freeze-thaw cycles. The PCR product has an A-tail at the 3’ end, allowing direct connection to T-vectors.

 

Components

Product NameProduct NumberSpecifications
2× Medino® Ultra-Long PCR Master MixLM201008-1-ES031 mL
LM201008-1-ES085 × 1 mL
LM201008-1-ES5050 × 1 mL
Components NumberComponents NameLM201008-1-ES03LM201008-1-ES08LM201008-1-ES50
LM201008-1-A2×Medino® Ultra-Long PCR Master Mix1 mL5 × 1 mL50 × 1 mL
LM201008-1-B25 mM MgSO4500 μL1 mL5 × 1 mL
LM201008-1-C10 mM dNTPs100 μL500 μL5 × 1 mL

Instructions

  1. Reaction System
ComponentsVolume (μL)
ddH2Oto 50 μL
2× Medino® Ultra-Long PCR Master Mixa)25 μL
Template DNAb)The appropriate amount
Forward primer (10 μM)2 μL
Reverse primer (10 μM)2 μL

[Note]:

  1. a) Reaction Buffer: The buffer contains 2.5 mM Mg2+ and optimized dNTPs. Additionally, the kit provides extra MgSO4 and dNTPs. For fragments larger than 20kb that are difficult to amplify, you can try adding 0.5-2 mM Mg2+ and 200-500 μM dNTPs.
  2. b) The recommended amounts of different templates are as follows:
Types of templatesRange of fragment amounts for a 50 μL reaction system
Genomic DNA or E. coli bacterial liquid10-1000 ng
Plasmid or viral DNA0.5-50 ng
cDNA1-5 μL (not exceeding 1/10 of the total PCR reaction volume)
  1. c) For amplifying long fragments, it is recommended to use a two-step method for annealing and extension. The annealing/extension time is set at 30 sec/kb, which can amplify most fragments. For fragments larger than 20 kb, the reaction time can be adjusted accordingly but should not exceed 60 sec/kb.
Cycle stepTemp.TimeCycles
Initial denaturation95 °C3 min1
Denaturation98 °C10 sec35
Annealing68 °C30 sec/kb
Final extension72 °C10 min1

Notes

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