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Dear Nenci I always used CIP with great success. A good control to test whether CIP worked, a plate with a plasmid (containing no insert) should not ligate and will not be circular, hence if CIP worked, this plate should not contain any plasmids grown on it (or very few). However, if you were to clone a gene, a plasmid that contains the inserted gene (which will bring in the two phosphates for the two dephosphorylated ends) will ligate and colonies will grow. I never tried SAP, thus cannot comment on that Good luck Akram Alian, Ph. D. Department of Biochemistry & Biophysics UCSF 600 16th street, S414 San Francisco, CA-94158 Phone: (415)-476-3937 From: CCP4 bulletin board [mailto:[log in to unmask]] On Behalf Of Nenci Simone Sent: Thursday, January 28, 2010 6:16 AM To: [log in to unmask] Subject: [ccp4bb] Phosphatase, which is the best? I need to dephosphorilate blunt ends and, surfing on internet I found this two enzymes: - Shrimp Alkaline Phosphatase (SAP) - Calf Intestinal Alkaline Phosphatase (CIP) Have you any experience about these phosphatases, I mean wich is the best for blunt ends? All the best Nenci Simone ------------------------------------------------- Dept. Genetics and Microbiology University of Pavia Via Ferrata 1, 27100 Pavia Italy Tel. +39-0382-985535/985534 Fax +39-0382-528496 Web www.unipv.it/biocry