Oct 09, 2022

Public workspaceTransformation

DOI
dx.doi.org/10.17504/protocols.io.36wgq7xpyvk5/v1
  • 1Tecnologico de Monterrey Campus Chihuahua
Denisse Chacón Ramírez
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DOI: 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,
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
Transformation
2h 30m 30s
2h 30m 30s
Mix Amount50 µL of fresh competent cells withAmount2 µL of genetic material (softly without pipetting) in a sterile 1.5 mL Eppendorf tube.

Incubate TemperatureOn ice for Duration00:30:00

30m
Apply thermal shock in a water bath at Temperature42 °C for Duration00:00:30

30s
Immediately, put TemperatureOn ice

AddAmount250 µL of SOC medium (tempered at Temperature37 °C ).

Incubate at Temperature37 °C forDuration01:00:00 at Shaker225 rpm .

1h
Disseminate Amount50 µL in a selective medium and incubate DurationOvernight at Temperature37 °C

1h