DH5Alpha Competency
The DH5Alpha cells were made competent on February 22, 2012 using the Inoue Protocol.
Contents
Section titled “Contents”1. Competency test procedure
Section titled “1. Competency test procedure”Greg Hild measured their competency by transforming the plasmid pUC19 into the cells at 5 ng, 500 pg, and 50 pg using the following procedure:
- Defrost the cells from -80C on ice (approximately 20 minutes).
- Add 50 uL of cells to an Eppendorf tube and 1 uL of the appropriate plasmid and flick the tube 6 times to mix.
- Ice for 5 minutes, heat shock in a 42C water bath for 42 seconds and then ice for 2 minutes.
- Add the 50 uL of cells to 500 uL of SOC and shake in the INNOVA shaker at 37C, 250 rpm for 1 hour.
- Plate 300 uL of cells and allow to grow in the incubator at 37 C for 24 hours.
2. Competency results
Section titled “2. Competency results”2.1 Competency 02-027-2013
Section titled “2.1 Competency 02-027-2013”- Third box in “Homemade cell” drawer
- Labeled DH5Alpha cells
2.2 Competency 03-30-2013
Section titled “2.2 Competency 03-30-2013”- First box in “Homemade cell” drawer
- Labeled “DH5Alpha cells (Matouschek Lab competency protocol)
2.3 Competency 05-13-2014
Section titled “2.3 Competency 05-13-2014”- first and second box in “Homemade cell” drawer
- Labeled DH5Alpha cells
