Plasmid DNA storage
Posted 06 January 2005 - 05:30 AM
planning to do pcr cloning of a promoter. will transfect the resulting reporter plasmid in cos cells. if i isolate the vector plasmid DNA from a megaprep. how long i can store it in the freezer. does long term storage cause any deterioration of the plasmid DNA.
Also please let me know the storage conditions.
Posted 06 January 2005 - 12:39 PM
Edited by green, 06 January 2005 - 12:40 PM.
Posted 08 January 2005 - 01:21 PM
Over-time, plasmids begin to deteriorate, but probaly not enough to prevent their use. If using old plasmids and you have trouble, you can run some on a gel (0.8-1.0% agarose), cut the band of supercoiled DNA, and purify to get rid of excess nicked DNA.
In addition to storing the plasmid, store a couple 1 ml aliquots of the transformed bacteria to innoculate culutres down the road. Add 300ul 50% glycerol to 700ul of your maxiprep culture and freeze. Try to use microtubes that have a rubber sealing ring in the lid. These bacterial stocks should be safe for years as well.
- The Biology Ninja
Edited by DointheScience, 08 January 2005 - 01:24 PM.
Posted 10 January 2005 - 05:26 AM
Posted 10 January 2005 - 09:00 AM