Primer

Part:BBa_K2973016:Experience

Designed by: Nick Delkis, Aphrodite Katsaouni   Group: iGEM19_Thessaly   (2019-09-28)


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Applications of BBa_K2973016

In our project, this primer (BBa_K2973016) was used along with its reverse primer (BBa_K2973017) for the amplification reactions of the IS6110 gene (BBa_K2973009) that was used as a biomarker, 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. 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

UNIQ1873a798aad801be-partinfo-00000006-QINU UNIQ1873a798aad801be-partinfo-00000007-QINU