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Team:Groningen/21 June 2010
From 2010.igem.org
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| - | [[Image:21-06-10-1gn.jpg|200px|thumb|left|Clones 1 and 2. + = cut with XhoI, P+ = cut with PstI and XhoI, S+ = cut with SpeI and XhoI, X+ = cut with XbaI and XhoI]] [[Image:21-06-10-2gn.jpg|200px|thumb|left| | + | [[Image:21-06-10-1gn.jpg|200px|thumb|left|Clones 1 and 2. + = cut with XhoI, P+ = cut with PstI and XhoI, S+ = cut with SpeI and XhoI, X+ = cut with XbaI and XhoI]] [[Image:21-06-10-2gn.jpg|200px|thumb|left|Clones 3 and 4.]] |
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Revision as of 14:09, 13 October 2010
Hydrophobofilm
pushing coatings into a greener future
Week 25, Arend Jan
Control restriction of pNZ8901-bbs with biobrick site enzymes. All were also cut with XhoI to check the orientation of the sites (in case the sites in the original plasmid were not removed).
- 10ul plasmid - 1.5ul buffer G (XbaI, SpeI) or R (PstI) - 0.5ul biobrick enzyme (XbaI, SpeI, or PstI) - 0.5ul XhoI - 2.5ul MQ
Because of the added XhoI a ~270bp fragment should be present. This is the case in all restrictions. All PstI restrictions give an unexpected 3 bands. The plasmid is ~800bp larger than the clone manager file but this was not considered a problem. It is likely that there is a PstI site in the unknown sequence. This site will have to be deleted before we can use this plasmid in multi-biobrick cloning steps.
As an extra control a restriction was done with BamHI which was used to linearize the plasmid after inserting the biobrick sites. Together with XhoI this should give a ~300bp band.
- 5ul plasmid - 1.5ul buffer G - 0.5ul BamHI - 0.5ul XhoI - 7.5ul MQ
Gel 23-06-10
