Glycerol Stocking

After successfully sequencing a bacterial plasmid, we must stock it for storage.

1. Add the plasmid to the database in Quartzy. See instructions on this page.

Important

Add plasmids to the database before stocking! That way, you don’t run into issues with overlapping numbering if someone else is also stocking around the same time.

2. Culture desired 4 mL of bacteria clone with the plasmid in LB Broth with the correct resistance antibiotic (overnight).

3. Add 2 mL of 60% glycerol to the 4 mL of culture. Mix well.

4. Aliquot 2 mL of the resulting mixture to three cryovials.

5. Label each cryovial with pKG#, date, initials, and description of the construct. Labels can be printed using the LabBot app on slack “Print Labels”. (If it does not show up begin to type “/labels” to find it.) You must be connected to MIT Wifi for this app to work.

6. Put cyrovials in -80 C. Put one cryovial per box.