|
Back to Protocols
SAP Dephosphorylation
Protocol for Dephosphorylation of 5'-ends of DNA:
1. Resuspend 1 pmol of DNA ends (about 1 μg of a 3 kb
plasmid) in nuclease-free water.
2. Prepare reaction mix in a 20 μl volume according to the
following:
- DNA - 1 μl
- 10X SAP Reaction Buffer - 2 μl
- Water, Nuclease-Free - up to 19 μl
- Shrimp Alkaline Phosphatase (1 unit/μl) - 1 μl
3. Incubate at 37°C for 30-60 min.
4. Stop reaction by heating at 65°C for 15 min. This
completely inactivates SAP.
Protocol according to USB www.usbweb.com
|
"
