Ligation
Ana Belem García González, Jair Alexis Gardea Sáenz, Irán Alessandra Chaparro Rodríguez, Georgina Elisa Diego Macías
Published: 2022-10-12 DOI: 10.17504/protocols.io.6qpvr4dmbgmk/v1
Abstract
Protocol for a Ligation with Master Mix Ligase after a restriction digestion protocol had been made.
Steps
1.
Add5µL of Master Mix Ligase
2.
Add the a quantity of the DNA vector digested by EcoRI and PstI in a molar ratio 1:3 with the DNA insert
3.
Add the a quantity of the DNA insert digested by EcoRI and PstI in a molar ratio 3:1 with the DNA vector
4.
Add water up to 20µL
5.
Ligate for 20h 0m 0s at 22°C
6.
Transform with5µLof the product
10.
10.
