Primer

Part:BBa_K2973017:Experience

Designed by: Nick Delkis, Aphrodite Katsaouni   Group: iGEM19_Thessaly   (2019-09-28)
Revision as of 20:05, 28 September 2019 by Nickdelkis (Talk | contribs) (→‎Applications of BBa_K2973017)


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Applications of BBa_K2973017

In our project, we used the IS6110 gene (BBa_K2973009) as a DNA template, in order to perform isothermal amplification reactions with Recombinase Polymerase Amplification (RPA). Using this method, we were able to detect quantities as low as 10 to the minus 9 nanograms in reaction times ranging from five to 20 minutes. The primers used in the amplification reactions were BBa_K2973016 (forward) and BBa_K2973017 (reverse). These primers include 5' overhangs ( a T7 promoter and a trigger sequence) to allow for transcription by a T7 RNA Polymerase and translation by a toehold-switch regulated system encoding for a reporter gene (BBa_K2973007). The expected amplified product length is 136 bp.

RPA reaction after clean up of the amplified product

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PCR reaction

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User Reviews

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