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Team:Groningen/21 June 2010
From 2010.igem.org
(New page: {{Team:Groningen/Header}} '''Week 25''' {{Team:Groningen/Footer}}) |
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{{Team:Groningen/Header}} | {{Team:Groningen/Header}} | ||
| - | '''Week 25''' | + | '''Week 25, Arend Jan''' |
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| + | 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). | ||
| + | |||
| + | |||
| + | <pre>- 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 </pre> | ||
| + | |||
| + | Gels 21-06-10 1+2 | ||
| + | |||
| + | |||
| + | 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. | ||
| + | |||
| + | <pre>- 5ul plasmid | ||
| + | - 1.5ul buffer G | ||
| + | - 0.5ul BamHI | ||
| + | - 0.5ul XhoI | ||
| + | - 7.5ul MQ </pre> | ||
| + | |||
| + | Gel 23-06-10 | ||
{{Team:Groningen/Footer}} | {{Team:Groningen/Footer}} | ||
Revision as of 14:00, 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
Gels 21-06-10 1+2
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
