3 replies to this topic

[serundipity2009]

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

[noelmathur]

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.

[Warren]

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..

[amelia417]

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.