Salmonella electrocompetent cells protocol

3/6/01 ECK
revised 8/6/2012

Description

Electrocompetent cells are normal cells suspended in a low conductivity medium which prevents arcing during electroporation. Always keep cells on ice, and drain pellets well after centrifugation! It is important to grow the cells to mid-log or earlier, especially for Salmonella .

• Resuspend pellet in 20 ml lowMho solution.
• 5' x 3,000 g in a refrigerated centrifuge.
• Discard the supernatant each time, including the last.

• Sit a metal block for Eppindorf vials on dry ice. Load it with 600 µL Eppindorf vials.
  
• Use electric pipetter to dispense 40 µL aliquots to the cold vials.
  
• Close vials and store at -70°. In this form, cells are good for several months.

lowMho solution
10% glycerol in deionized distilled H2O Sterilize in autoclave. Last Update: Thursday June 19 2014 This page has been viewed Failed to execute CGI : Win32 Error Code = 2
times. Eric Kofoid eckofoid at ucdavis.edu