Team:Groningen/5 July 2010
From 2010.igem.org
Week 27, Arend Jan
The pMASK-EH construct is cut with EcoRI and SpeI to get the fragments containing the ChpE and ChpH genes and ligate them in to the expression plasmid. A mix of both genes will be used for ligation and the clones will be checked afterwards for which gene they contain.
- 10ul plasmid pNZ8901-bbs - 1.5ul buffer Tango - 0.5ul SpeI - 3ul MQ - perform digestion - add 1.88ul buffer Tango - add 0.5ul EcoRI - 17.5ul plasmid pMASK-EH - 2ul buffer Tango - 0.5ul SpeI - perform digestion - add 2.5ul buffer Tango - add 0.5ul EcoRI
Ligation of ChpE and ChpH into pNZ8901-bbs:
- 2/5ul backbone - 14/11ul insert (ChpE+H) - 2ul T4 buffer - 2ul T4 ligase
Colonies were picked, grown, and prepped for restriction checks (next week).