Difference between revisions of "Part:BBa K2686006:Experience"
 
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The part was designed and constructed in pET vector. The sequence was then confirmed on pET vector using sanger sequencing. The part was expressed in the pET vector using a BL21 DE3 cell free expression system. After expression and purification, the part was validated using a native gel, with multiple positive and negative controls (review iGEM EPFL 2018 results page).
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The part was designed and constructed in the original pET14 vector from <bbpart>K2686001</bbpart>. The sequence was confirmed using Sanger sequencing. The part was expressed in the pET14 vector using a BL21 DE3 TX-TL (Sun et al., 2013) cell free expression system. After expression and purification, the part was validated in multiple ways. First, multiple SDS-PAGE experiments indicate that our expressed protein is at the correct hypothetical size, for the 60-mer and that sfGFP binds to the encapsulin.
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The part was then amplified out of the pET vector and inserted into the pSB1C3 plasmid backbone, and submitted. There was not enough time to perform Sanger sequencing, but colony PCR gave us the correct insert size.
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Latest revision as of 12:49, 17 October 2018

The part was designed and constructed in the original pET14 vector from K2686001. The sequence was confirmed using Sanger sequencing. The part was expressed in the pET14 vector using a BL21 DE3 TX-TL (Sun et al., 2013) cell free expression system. After expression and purification, the part was validated in multiple ways. First, multiple SDS-PAGE experiments indicate that our expressed protein is at the correct hypothetical size, for the 60-mer and that sfGFP binds to the encapsulin.


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UNIQcf142b1fb048abed-partinfo-00000001-QINU UNIQcf142b1fb048abed-partinfo-00000002-QINU