Team:SDU-Denmark/Protocols/Restrictions
From 2009.igem.org
Sadly, we had a lot of trouble with our restrictions, and this page will reflect that in including alot of different restriction protocols.
Protocol 1
- Pool plasmid and dry on vacuum centrifuge down to about 50uL
- Mix the following into one tube:
- 4uL Plasmid and RIP
- 5uL 10x Buffer
- 0,5uL BSA
- Fill with water to 47uL (37,5uL)
- 1,5uL Enzyme 1
- 1,5uL Enzyme 2
- Incubate for 2 hours on 37 degress C.
- Inactivate the enzymes at 80 degress C for 20 minutes.
- Place product on -20 degress C or continue immediately to ligation.