Part:BBa_K2246000:Experience

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Applications of BBa_K2246000

Confirmation of the Blue color in the colonies. The blue chromoprotein expression of the BSLA construction (Blue chromoprotein, sense siRNA, loop and anti-sense siRNA) was confirmed in 2 strains of Escherichia coli: HT115 and DH5α.

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Figure 1 and 2: Blue chromoprotein expresion of the BSLA-AWD device in DH5α (left) and HT115 (right).

SiRNA production confirmation The siRNA confirmation protocol was made to test, with the real time PCR, the presence of the siRNAs designed by the iGEM Tec Cem team. The four devices capable of producing the siRNAs were tested: BSLA-SOD, BSLA-WNT, BSLA-RACI and BSLA -AWD. The protocol used for this purpose is based on the liquid northern hybridization proposed by Wang X., Tong Y. and Wang S. (2010). The protocol is described as a novel form to detect miRNAs. We expect to detect the siRNAs that our different constructs can produce taking advantage of their hybridization with the forward and reverse templates of their DNA sequences and the ability of the exonuclease to digest the non-hybridized RNA sequences.

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Figure 2. RNA from the HTT115 strain witouth the siRNA construct and with the siRNA construct (BSLA-AWD) with different hybridization oligos and treatments in a 12% no-denaturing gel at 100 Volts.

User Reviews

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