DNA extractio kit - (May/03/2011 )
Hi guys,
I need to know whats the diff. b/w extracting DNA from kits and by normal lab protocols...time? efficiency? relative cost?
I know its a general questions but I haven't used it before. Please help me.
Thanks
mostly time ...since by using e.g. silica membranes you can skip the precipitation step (that you need for most lab protocols) and you can directly from the membrane with water.
for plasmid isolations you can use the alkaline lysis method (described by Birnboim and Doly) ...this is also fast and cheap ...and gives good quality.
Regards,
p
pDNA on Wed May 4 10:22:36 2011 said:
mostly time ...since by using e.g. silica membranes you can skip the precipitation step (that you need for most lab protocols) and you can directly from the membrane with water.
for plasmid isolations you can use the alkaline lysis method (described by Birnboim and Doly) ...this is also fast and cheap ...and gives good quality.
Regards,
p
usually for lab protocols it will take the entire day. so how much of time will be reduced if I use the kit. what are the steps involved?
Dear Indus,
it would be much easier if you could specify what DNA extraction you wanna perform? ...plasmid DNA isolation? ...genomic DNA isolation? ...and from what source? Bacteria? Eukaryota?
Regards,
p
InduS on Wed May 4 13:04:04 2011 said:
pDNA on Wed May 4 10:22:36 2011 said:
mostly time ...since by using e.g. silica membranes you can skip the precipitation step (that you need for most lab protocols) and you can directly from the membrane with water.
for plasmid isolations you can use the alkaline lysis method (described by Birnboim and Doly) ...this is also fast and cheap ...and gives good quality.
Regards,
p
usually for lab protocols it will take the entire day. so how much of time will be reduced if I use the kit. what are the steps involved?
Plus, it's usually more expensive to use a kit rather than preparing your own reagents.
using kits reduces the variability of the isolations by providing standardized buffers and other materials. lot to lot variations are minimized.
so, even if the kit costs a little more than preparing your own you save time and related expenses caused by variability with homemade.
I did E. coli DNA extraction using alkaline lysis, and it's good for quick screen, digestion. But when it comes to sequencing ..... wasted so much time cleaning that DNA and repeating sequencing reactions, it would be a lot cheaper with a kit.
gDNA (played only with plants) would never do without a kit
I clean my alkaline lysis up following the protocol for PEG precipitation in the ABI guide. It works just fine.
I clean my Alk lysis minipreps up following the protocol for PEG precipitation in the ABI guide. It works just fine, takes a while to process a whole lot, but it's not too bad.