Can I store plasmid in 4C refrigerator? - (Jun/09/2008 )
I just received a small amount of plasmid dissolved in pure H2O. It came by air mail, and was not refrigerated or anything. I put it into our 4C fridge. Is that ok for one or two weeks?
-brightfield-
I think it would be better if it is frozen down for long-term storage. It might get degraded (partially).
-zek-
QUOTE (brightfield @ Jun 9 2008, 10:43 AM)
I just received a small amount of plasmid dissolved in pure H2O. It came by air mail, and was not refrigerated or anything. I put it into our 4C fridge. Is that ok for one or two weeks?
Yep.
Partial degradation and de-supercoiling may occur, but should be good for most of the mol bio purposes, especially if you are going to make a large batch using this plasmid.
-cellcounter-
QUOTE (cellcounter @ Jun 9 2008, 03:58 PM)
QUOTE (brightfield @ Jun 9 2008, 10:43 AM)
I just received a small amount of plasmid dissolved in pure H2O. It came by air mail, and was not refrigerated or anything. I put it into our 4C fridge. Is that ok for one or two weeks?
Yep.
Partial degradation and de-supercoiling may occur, but should be good for most of the mol bio purposes, especially if you are going to make a large batch using this plasmid.
Yep, we are going to do a MaxiPrep on it, hopefully, within the next couple of weeks. Actually, I learned the plasmid is probably in a TE buffer. I assume that can only help, right?
-brightfield-
Why not just stick it in the -20? There's no reason to leave DNA in the 4C; it takes just minutes to thaw.
-Judes-
TE at 4C is fine, but I'd still try to get that maxiprep done quickly.
-swanny-
QUOTE (Judes @ Jun 10 2008, 11:53 AM)
Why not just stick it in the -20? There's no reason to leave DNA in the 4C; it takes just minutes to thaw.
Agreed. I can only see this as laziness.
-killerkoz17-
QUOTE (killerkoz17 @ Jun 9 2008, 11:41 PM)
QUOTE (Judes @ Jun 10 2008, 11:53 AM)
Why not just stick it in the -20? There's no reason to leave DNA in the 4C; it takes just minutes to thaw.
Agreed. I can only see this as laziness.
Not at all, the -20 is right next to the 4C. However, I thought it is generally not good to go through freeze-thaw cycles if not necessary.
-brightfield-
QUOTE (brightfield @ Jun 11 2008, 07:55 PM)
QUOTE (killerkoz17 @ Jun 9 2008, 11:41 PM)
QUOTE (Judes @ Jun 10 2008, 11:53 AM)
Why not just stick it in the -20? There's no reason to leave DNA in the 4C; it takes just minutes to thaw.
Agreed. I can only see this as laziness.
Not at all, the -20 is right next to the 4C. However, I thought it is generally not good to go through freeze-thaw cycles if not necessary.
You can aliquote your DNA. I do that usually for plasmids which I know I am gonna use often.
-zek-
QUOTE (brightfield @ Jun 12 2008, 03:55 AM)
QUOTE (killerkoz17 @ Jun 9 2008, 11:41 PM)
QUOTE (Judes @ Jun 10 2008, 11:53 AM)
Why not just stick it in the -20? There's no reason to leave DNA in the 4C; it takes just minutes to thaw.
Agreed. I can only see this as laziness.
Not at all, the -20 is right next to the 4C. However, I thought it is generally not good to go through freeze-thaw cycles if not necessary.
Oh well, yeah, true. But it's still better to store it in the -20C. The DNA will be fine at 4C, i guess if you're using it regularly in a few day period then ok. But keeping it in there for a long time is not the best way to store it. Enzymes that degrade the DNA will have some activity at 4C, whereas -20C they'll have none. Water crystal shearing is the only real problem in -20C but this really is nothing compared to enzyme degradation, i don't know if water crystals even do much to DNA but nucleases at 4C certainly will.
The other thing is that your solution will evaporate at 4C eventually, not a major issue, just resuspend, but -20C is better.
-killerkoz17-