Knight:TOPO vector preparation
This is a made-up protocol to prepare pSB**5 vectors for TOPO cloning. It has not been verified at all!
- X μL plasmid
- 1 μL Nt.BstNB I
- 5 μL NEBuffer 3
- 44-X μL H2O
- Incubate at 55°C for at least 1 hour
- Heat-inactivate at 80°C for 20 mins
Do a purification step? Try the same buffer?
Covalently attach DNA topoisomerase I from Vaccinia
Quality control procedure
Unit test definition
Alternative protocol from Shuman et al.
From Shuman 1994: maximum topoisomerase activity was observed with a 20 μL reaction mixture containing
Do a purification step? Or use it directly?