Oct 11, 2022
- Georgina Elisa Diego Macías1,
- Ana Belem García González1,
- Irán Alessandra Chaparro Rodríguez1,
- Jair Alexis Gardea Sáenz1
- 1Tecnologico de Monterrey Campus Chihuahua
Protocol Citation: Georgina Elisa Diego Macías, Ana Belem García González, Irán Alessandra Chaparro Rodríguez, Jair Alexis Gardea Sáenz 2022. Ligation. protocols.io https://dx.doi.org/10.17504/protocols.io.6qpvr4dmbgmk/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: October 11, 2022
Last Modified: October 11, 2022
Protocol Integer ID: 71134
Abstract
Protocol for a Ligation with Master Mix Ligase after a restriction digestion protocol had been made.
Materials
- Ultra Pure Water
- Digested fragment of DNA vector with EcoRI & PstI
- Digested fragment of DNA insert with Xbal Pstl
- Master Mix Ligase
Add5 µL of Master Mix Ligase
Add the a quantity of the DNA vector digested by EcoRI and PstI in a molar ratio 1:3 with the DNA insert
Add the a quantity of the DNA insert digested by EcoRI and PstI in a molar ratio 3:1 with the DNA vector
Add water up to 20 µL
Ligate for 20:00:00 at 22°C
20h
Transform with5 µL of the product