DNA
I-SceI RS
Part:BBa_K175027:Experience
Designed by: Tim Weenink Group: iGEM09_TUDelft (2009-08-01)
Revision as of 04:13, 1 November 2009 by Tim Weenink (Talk | contribs) (→Applications of BBa_K175027)
Sequence has been shown to be cut by I-SceI in vitro.
Characterization of BBa_K175027
Gel electrophoresis of BBa_K175027 in pSB1AK3 backbone. Lane 1: undigested (supercoiled) plasmid. Lane 2: PstI digested part, expected size = 5095bp. Lane 3: PstI and I-SceI restricted part, expected band lenghts are 3204 and 1891bp. Expected lenghts correspond to the bands observed in the gel.
This biobrick was successfully sequenced and tested for functionality. Sequencing results can be found on the part page. Functionality was assessed using commercially available I-SceI restriciton enzyme. The plasmid was purified and subjected to a digestion with BglI alone and BglI plus I-SceI. The latter would theoretically result in two fragments of 2275 and 944bp.
The results in the adjecent image show that the digestion indeed results in the expected band pattern. This shows that the restriction site works as expected.
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