Feb 21, 2022
Version 2
Transformation Protocol V.2
- 1New England Biolabs
- New England Biolabs (NEB)Tech. support phone: +1(800)632-7799 email: info@neb.com
Protocol Citation: New England Biolabs 2022. Transformation Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.bd2yi8fwVersion created by New England Biolabs
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: March 21, 2020
Last Modified: February 21, 2022
Protocol Integer ID: 34616
Keywords: quick ligation transformation, comp cells, Transformation, Competent cells
Abstract
Quick Ligation products may be transformed by many different methods. The following protocol is recommended by New England Biolabs.
Safety warnings
Please refer to the Safety Data Sheets (SDS) for health and environmental hazards.
Thaw competent cells On ice .
Chill approximately 5 ng ligation mixture (2 µL ) in a 1.5 ml microcentrifuge tube.
Add 50 µL competent cells to the DNA.
Mix gently by pipetting up and down or flicking the tube 4–5 times to mix the cells and DNA. Do not vortex.
Place the mixture On ice for 00:30:00 . Do not mix.
Heat shock at 42 °C for 00:00:30 . Do not mix.
Note
Please note: For the duration and temperature of the heat shock step as well as for the media to be used during the recovery period, please follow the recommendations provided by the competent cells’ manufacturer.
Add 950 µL room temperature media to the tube.
Place tube at 37 °C for 01:00:00 . Shake vigorously (250 rpm ) or rotate.
Warm selection plates to 37 °C .
Spread 50 µL –100 µL cells and ligation mixture onto the plates.
Incubate Overnight at 37 °C .