PROTOCOL 002: GLYCEROL STOCK OF PLASMID-CONTAINING BACTERIA

 

REAGENTS

 

Bacterial culture

Sterile Glycerol

Screw-cap 1.5 mL tube

Labeling Tape

 

PROCEDURE

 

**NOTE** All procedures should be performed aseptically.

 

Cut a length of lab tape slightly longer than the circumference of the screw-cap tube.

 

Write the following information on the label:

 

Plasmid Name

Antibiotic resistance: Amp, Kan, Tet, etc./Strain of E. coli (DH5a, Top10F, etc).

Date/Initials

 

Place label around screw-cap tube.

 

Transfer 850 mL from culture flask to screw-cap tube.

 

Add 150 mL of glycerol.

 

Carefully pipette solution to ensure thorough mixing.

 

Place in -80 C freezer in appropriate glycerol stock box.

 

Enter Rack, Box and Slot information into Plasmid Database.