DNA Ligation Calculator

Calculate optimal DNA ligation conditions including insert-to-vector ratios, DNA amounts, and ligase concentrations for molecular cloning.

DNA Parameters

bp

Length of vector DNA in base pairs

bp

Length of insert DNA in base pairs

Vector DNA concentration

Insert DNA concentration

Molar ratio of insert to vector

µL

Total ligation reaction volume

Ligation Results

Enter DNA parameters to calculate ligation conditions

DNA Ligation Guidelines

Optimal Conditions

  • • Insert-to-vector ratio: 2:1 to 5:1
  • • Total DNA: 50-200 ng per reaction
  • • T4 DNA Ligase: 1-5 units
  • • Incubation: 16°C overnight or RT 1-4 hours

Tips for Success

  • • Use fresh, high-quality DNA
  • • Ensure compatible sticky ends
  • • Include appropriate controls
  • • Optimize ratio based on insert size