Mini-Prep Blues

Soeren Jensby Nielsen sjn at biobase.aau.dk
Tue Jul 12 06:23:44 EST 1994


In <Csor2t.KFx at usenet.ucs.indiana.edu> jgraham at bronze.ucs.indiana.edu (the End) writes:

>You may want to try alkaline lysis again, but use ammonium acetate as your 
>third solution rather than sodium or potassium acetate*. For cells
>resuspended in 200 ul and denatured with 400 ul of SDS/NaOH, add
>300 ul of 7.5 ammonium acetate at the dissolved pH. This change 
>dramatically increase the number of enzymes which cut well in my
>experience.

>For super-clean sequencing templates, add RNAase to the supernatant
>follwing this step, precipitate with 400 ul of isopropanol, and then 
>resuspend the pellet in 2 M ammonium acetate, spin out the  "rocks"
>and reprecipitate**
>Jim
>J. E. Graham 
>* Morelle, G. Focus 11:1 p.7
>** Lee and Rasheed. 1990. Biotechniques 9(6) p.676 


Jim is right. The protocol by Lee & Rasheed is the best.


Soren.




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