7 replies to this topic

[porawa]

I fail to extract DNA plasmid by Eppendrof.

[thegradstudent]

try QIAGEN

[marc_U7snurp]

Had the same problem with an eppendorf kit, well I asked another one and It worked.

Did you add the appropriate antibiotic to the culture? If not the plasmid will not stay in the bacteria and you'll get no plasmid.

If its not this, then maybe call them and ask for hints andf help.

Cheerz,

marc

[fred_33]

hi
i use qiagen column kits for my plasmid extraction and it is ok.
If the plasmid i want to extract is a low-copy, i increase the volume of bacteria culture (up to 200ml) with adapting protocol according to volume of culture.

Fred

In tis topic, i give the protocol i use now, cheaper. I like this one because i can easily adapt the quantity if i want more plasmid.
http://www.protocol-online.org/forums/inde...t=0&#entry11777

Edited by fred_33, 04 March 2005 - 02:40 AM.

[Gerd]

thegradstudent, on Feb 22 2005, 07:25 AM, said:

try QIAGEN

Qiagen most often gives clean DNA, but not much of it.
I liked the BioRad kit - good yield, saves a lot of work and money when you need a little more of your plasmid.

[methylnick]

porawa, on Feb 21 2005, 01:49 PM, said:

I fail to extract DNA plasmid by Eppendrof.

Are you sure the strain of bacteria you are using is compatible with the kit? I remeber some strains are not optimal using this kit.

Nick

[postdoc2130]

We and others have good experience with GenCatch plasmid DNA prep kits.
High yield, high purity (can be sequenced upto ~900 bp signgle round), easy, fast and much cheaper!

[badcell]

Macherey-Nagel is the same as Qiagen but cheaper. I agree with Gerd that Qiagen (and Macherey-Nagel) gives not much DNA (depending on the plasmid), but the DNA you get is quite clean.