Jan 13, 2026
Transformation Protocol
- Tamra TCL Lahom1
- 1University of Pennsylvania
Protocol Citation: Tamra TCL Lahom 2026. Transformation Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.5jyl8xpr6v2w/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: January 12, 2026
Last Modified: January 13, 2026
Protocol Integer ID: 238476
Keywords: transformation protocol quick ligation product, following protocol, protocol, new england biolab
Abstract
Quick Ligation products may be transformed by many different methods. The following protocol is recommended by New England Biolabs.
Troubleshooting
Safety warnings
* 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.
Before start
Make selection plates prior to starting this procedure.
Protocol
3h 0m 30s
Thaw competent cells on ice for 00:30:00 .
30m
Take the concentration of the plasmid sample. It was 10 mg/mL
Add 10 ng (2 µL µl) of the plasmid to 50 µL of competent cells. Gently flick the tube 4–5 times to mix the cells and DNA. Do not vortex.
Chill approximately 5 ng (2 µL µl) of the plasmid in a 1.5 ml microcentrifuge tube.
Place the mixture on ice for 00:30:00 minutes. Do not mix.
30m
Heat shock at 42°C for 00:00:30 . Do not mix.
30s
Add 300 µL µl of room temperature LB media to the tube.
Place tube at 37 °C for 02:00:00 minutes. Shake at 250 rpm.
2h
Warm selection plates to 37 °C
Spread 150 µL of the cells and ligation mixture onto the plates.
Incubate overnight at 37 °C .