short protocols in molecular biology...... - maybe too short??? (Jun/17/2006 )

Hi,
We use a lot of kits, for almost everything. We saw that in spite of they are expensive, at the end, we save money.
I used to do the whole protocol for my minipreps (or use kit). Very often I have to do 100 minipreps or more per day (in addition of a lot of more work!) it was horrible and expensive !!!!!!
So, now, for 1.5 ml-culture, I just add 100 ul sol I, mix, 200 ul sol II mix by inverting (no incubation), 150 ul sol III mix by inverting (incubate 1 or 2 minutes), centrifuge for 5 min. Precipitate with 0.7 V isopropanol. Centrifuge 5 min, wash with 70% ethanol. And elute in water with or without RNAsa. (I have a big stock of water with RNAsa 20 ug/ml final concentration). So I finish very quickly! And believe me. I digest with almost any restriction enzyme, and I have never had problems. Some times, I use those digestions for cloning without any problem (digest, ligate, tranform).
Ah, when I’m digesting just for screening colonies, I use re-used tips for the preps!!
I use kit just when I have to sequence, or transfect (endo-free kit) or something like that!

I think you should try ones to see if it works for you too.

Hi,
We use a lot of kits, for almost everything. We saw that in spite of they are expensive, at the end, we save money.
I used to do the whole protocol for my minipreps (or use kit). Very often I have to do 100 minipreps or more per day (in addition of a lot of more work!) it was horrible and expensive !!!!!!
So, now, for 1.5 ml-culture, I just add 100 ul sol I, mix, 200 ul sol II mix by inverting (no incubation), 150 ul sol III mix by inverting (incubate 1 or 2 minutes), centrifuge for 5 min. Precipitate with 0.7 V isopropanol. Centrifuge 5 min, wash with 70% ethanol. And elute in water with or without RNAsa. (I have a big stock of water with RNAsa 20 ug/ml final concentration). So I finish very quickly! And believe me. I digest with almost any restriction enzyme, and I have never had problems. Some times, I use those digestions for cloning without any problem (digest, ligate, tranform).
Ah, when I’m digesting just for screening colonies, I use re-used tips for the preps!!
I use kit just when I have to sequence, or transfect (endo-free kit) or something like that!

I think you should try ones to see if it works for you too.

i use qiaprep spin miniprep kit from qiagen. and it works like like a shot every time. highly reprodicible, fast, and not in the expensive bracket at all......even for gel purifying dna i prefer the qiagen min-elute kit rather than taking the lmp agarose and hot phenol way.....

- viv

If you've see my other posts i try not to plug, but...

There are kits and manufacturers out there that provide tools for most common molecular and cell bio techniques, that work just as effectively as the big box manufacturers like Promega and Qiagen. Their kits can be just as reliable and just as everyone here has said, you will really save a lot of time.

The smaller manufacturers can be harder to find, because they don't spend their ad budgets on a mass audience like Eppendorf.

But, you can start with our website www.vasus.com. We continue to add new manufacturers everyday. Though some larger manufacturers are listed with us, we really try to bring a wider diversity of products from less known manufacturers with better terms and pricing for the researcher.

Discount pricing, up to 70% off, is available without registration. If you need some supplier suggestions let me know.

mean this one?

Promega Wizard Miniprep Kit