low copy number plasmid purification kit - (Sep/12/2018 )
i am trying to purify my plasmid.
my plasmid is a low copy number.
my cells are XL 1 blue cells.
i used Hispeed Midi purification kit and i ended with a low yield.
from your experiences, if i want to end with approx. 1000ng/ul, which kit shall i use ??
Qiagen Hispeed Maxi ?? or do you prefer others ??
Our lab used Qiagen kits, either the QIAfilter maxi or the Large Construct kit (which is for plasmids or BACs that are both large and low-copy
Obviously you will get lower yield than you are used to with high copy plasmids. If the quality of the prep is good, just low yield, you can make several preps and pool them, then concentrate them by ethanol precipitation to get the desired concentration. Or, stepping up from midi to maxi should help.
thank you OldCloner!
yes i followed the LC instructions, but still i got low yields.
But, the kit already stated that with LC plasmids expect considerably low yields!
i did so, but ended having much to deal with, so i thought if i scale up to the maxi, things will be easier,
even if i pooled, at least each single yield would be higher than my previous.
thank you again!
Yeah, maxiprep for low-copy plasmid purification with required 1ug/ul concentrations would probably be a better choice.
Last time, after failing to grow with chloramphenicol, we used a reagent by Epicenter. I dont remember its name but it really helped us. We also used 300+ rpm and longer incubation.