Oct 09, 2022

Transformation

DOI
https://dx.doi.org/10.17504/protocols.io.36wgq7xpyvk5/v1
Transformation
  • 1Tecnologico de Monterrey Campus Chihuahua
Denisse Chacón Ramírez
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DOI: https://dx.doi.org/10.17504/protocols.io.36wgq7xpyvk5/v1
Protocol Citation: Ana Belem García González, Georgina Diego, Irán Alessandra Chaparro Rodríguez, Jair Alexis Gardea Sáenz 2022. Transformation. protocols.io https://dx.doi.org/10.17504/protocols.io.36wgq7xpyvk5/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: August 08, 2022
Last Modified: October 09, 2022
Protocol Integer ID: 68366
Keywords: Transformation, E. coli, transformation, cell
Abstract
A protocol for transforming E. coli cells is shown.
Materials
MATERIALS:
  • Competent cells
  • LB plates + antibiotic (depends on the used part)
  • Drigalski spatula
  • Eppendorf tubes of 1.5 mL


SOLUTIONS:
  • SOC medium


EQUIPMENT:
  • Water Bath at 42 °C
  • Cooler and ice
Transformation
2h 30m 30s
Mix 50 µL of fresh competent cells with2 µL of genetic material (softly without pipetting) in a sterile 1.5 mL Eppendorf tube.

Incubate On ice for 00:30:00

30m
Apply thermal shock in a water bath at 42 °C for 00:00:30

30s
Immediately, put On ice

Add250 µL of SOC medium (tempered at 37 °C ).

Incubate at 37 °C for01:00:00 at 225 rpm .

1h
Disseminate 50 µL in a selective medium and incubate Overnight at 37 °C

1h