Protocol Online logo
Top : New Forum Archives (2009-): : Molecular Cloning

Miniprep - (Nov/14/2009 )

Hello ALL,

For miniprep, the recommened incubation time is 12-16 hours and i was wondering if anyone has tried incubating for less time, such as 5-8 hours? Does less incubation time still give DNA?

THanks :lol:

-serundipity2009-

AFAIK, it also depends on copy number of your plasmid in said bacteria. If your plasmid is low-copy number, I would not mind going for 24 hours. I have done 8 hours sometimes but I knew that my plasmid is high copy number and I got enough DNA.

If you don't know give a try and see what results you get. RUn 2 cultures side by side.

-noelmathur-

noelmathur on Nov 14 2009, 01:14 PM said:

AFAIK, it also depends on copy number of your plasmid in said bacteria. If your plasmid is low-copy number, I would not mind going for 24 hours. I have done 8 hours sometimes but I knew that my plasmid is high copy number and I got enough DNA.

If you don't know give a try and see what results you get. RUn 2 cultures side by side.


if its high copy and the culture is turbid, you can get DNA after only 6 hours, it just will be a lower amount....I have done this in the past when trying to get things done FAST....warren..

-Warren-

serundipity2009 on Nov 14 2009, 11:13 AM said:

Hello ALL,

For miniprep, the recommened incubation time is 12-16 hours and i was wondering if anyone has tried incubating for less time, such as 5-8 hours? Does less incubation time still give DNA?

THanks :)

I am using several high copy plasmids and grow the mini prep bacteria for 8 hours from a single colony. Any less time and there is a pellet, it is just smaller and you will of course end of with less DNA in your mini. We only do the 12-16 hr incubation for maxi preps.

-amelia417-