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Qiagen mini-prep kit


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#1 Samantha

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Posted 07 February 2009 - 01:53 AM

Hi!

I prepared 4 tubes with inoculating bacteria. Half of them were purified by Qiagen mini-prep kit and the other half were purified by another kit (with P1+P2+P3 and quick column, no isopropanol precipitation reqquired). The yield and the quality prepared by the latter kit was fine, but i couldnt prep my plasmid by the QIagen kit. Has anyone also entercountered problems with the Qiagen kit? Thanks

Samantha

#2 scolix

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Posted 07 February 2009 - 03:21 AM

Hi!

I prepared 4 tubes with inoculating bacteria. Half of them were purified by Qiagen mini-prep kit and the other half were purified by another kit (with P1+P2+P3 and quick column, no isopropanol precipitation reqquired). The yield and the quality prepared by the latter kit was fine, but i couldnt prep my plasmid by the QIagen kit. Has anyone also entercountered problems with the Qiagen kit? Thanks

Samantha



Usually the Qiagen columns are fine. Rarely one or two columns turn out bad and one loses lots of DNA. But again its rare.

#3 Samantha

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Posted 09 February 2009 - 04:56 AM

Sorry to mention that the Qiagen mini-prep i used is the one makes use of gravity flow and isopropanol precipitation, not the quick spin column one.

Samantha

#4 94mkiv

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Posted 09 February 2009 - 10:00 PM

I haven't used the Qiagen mini prep kit using the gravity flow but I'm assuming it's similar to the gravity flow using the maxi kit.

When you say you" couldn't prep your plasmid using the Qiagen kit" do you mean you got no DNA from the prep?

Were the P1+P2+P3 used for each prep the same or was one from the Qiagen kit and another from the other kit?

I remember getting very few bad kits from Qiagen. It's rare but possible.

Did you ensure the P2 had no ppt and was still good. I remember getting no yields when someone hadn't closed the P2 bottle tightly.


-Mike

#5 Samantha

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Posted 09 February 2009 - 10:59 PM

I haven't used the Qiagen mini prep kit using the gravity flow but I'm assuming it's similar to the gravity flow using the maxi kit.

When you say you" couldn't prep your plasmid using the Qiagen kit" do you mean you got no DNA from the prep?
--> the yield was very very low...eluted in 30ul dH2O with only about 10ng/ul

Were the P1+P2+P3 used for each prep the same or was one from the Qiagen kit and another from the other kit?
--> i dont know if the P1+P2+P3 are the the same for the two kits...but the other kit uses a quick spin column instead of gravity flow

I remember getting very few bad kits from Qiagen. It's rare but possible.

Did you ensure the P2 had no ppt and was still good. I remember getting no yields when someone hadn't closed the P2 bottle tightly.
-->yes, no ppt at all...


-Mike






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