Difference between revisions of "Part:BBa K1636002"
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| + | The linker was cloned into plasmid pSB1C3, previously cut with EcoRI and PstI, and then a purification was performed. Several clones were selected to isolate plasmid by miniprep (figure 1A). In order to observe the integrity of the four extracted plasmids an electrophoresis was run (figure 1B). 10 microliters were loaded into each lane of plasmid, loaded with 4 microliters of buffer. | ||
Revision as of 01:07, 19 September 2015
Linker
This linker was designed by the iGEM TecCEM team in order to retrieve single-stranded oligonucleotides when produced as single-stranded genetic material (e.g. plasmid). The linker should be flanking said oligonucleotides, and given the design of its sequence, it is able to form a hairpin-like secondary structure that allows the usage of a restriction enzyme (MlyI from New England BIolabs) with a blunt cut thereby releasing the oligos of interest.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
The linker was cloned into plasmid pSB1C3, previously cut with EcoRI and PstI, and then a purification was performed. Several clones were selected to isolate plasmid by miniprep (figure 1A). In order to observe the integrity of the four extracted plasmids an electrophoresis was run (figure 1B). 10 microliters were loaded into each lane of plasmid, loaded with 4 microliters of buffer.