Suggestion needed for miniprep - (Jun/16/2005 )
Which kits are better for DNA miniprep
Qiagen miniprep kits
Eppendorf FastPlasmid Mini Kits?
The cheaper one!!!
I've tried both and found similar yields and didn't find any problem sequencing, ligating or performing downstream applications with either.
The eppendorf kit is certainly a bit quicker than the qiagen one but is only compatibile with a certain number of cell lines, you should check which ones. They work fine with DH5-alphas
I am using Qiagen miniprep kits. It works well for me.
i vote Qiagen
I never realised the Eppendorf kit is only good for certain cell lines, I tried it in Dh5a, XL1blue and Top 10s without anyproblem. Something I hadn't considered and will have to keep my eyes open for when trying kits.
Thanks for the tip.
Why use a kit for miniprep purposes in the first place?
Sambrooks alkali lysis method is just as easy and far cheaper.
yeah Eppendorf uses a lysozyme mediated step which is not amenable for certain cell lines, I tested the eppy kit before but have stuck with promega's kit.
you are absoultely correct, all these kits are based on the alkali lysis from the Manni bible. It's a right real shame that students these days only know of kits and use them routinely without knowing the background, the basics.
I guess the one advantage kits have is their QC. Generally speaking you know the quality of DNA you are going to get out of the kit, whereas sometimes quality can be an issue in the manual way.
Back when I was doing my Undergraduate honours year my supervisor would not supply me with kits so had to do things the old fashioned, including alkaline lysis for mini preps and Cs-Cl gradients for larger production and Cacium Phosphate transfections (although did later more on to lipid transfections). I have thanked him for this many times, as developed trouble shooting skills and an appreciation of process maybe all students should be banned from using kits. Although for the rest of us I'd like to avoid the Cs-Cl gradients if possible.
Yah, Scott, I can dig the whole thing with QC and all.
But the thing is this one... most of the time you do a mini prep to screen for recombinants. Sambrooks method is perfect for that as people generally screen by restriction digest. Which is perfectly compatable with Sambrooks method.
So great, now you want to sequence, transfect, whatever. choose one or two clones, put them through the kit protocol... just starting with plasmid or a mini/midi prep and put it though the kit protocol. Saves on a lot of expensive kit reagents.