Difference between revisions of "Part:BBa K2973009:Experience"
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===Applications of BBa_K2973009=== | ===Applications of BBa_K2973009=== | ||
| − | 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. | + | In our project, we used the IS6110 gene (<partinfo>BBa_K2973009</partinfo>) 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 <partinfo>BBa_K2973016</partinfo> (forward) and <partinfo>BBa_K2973017</partinfo> (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 (<partinfo>BBa_K2973007</partinfo>). The expected amplified product length is 136 bp. |
RPA reaction after clean up of the amplified product: | RPA reaction after clean up of the amplified product: | ||
Latest revision as of 12:55, 21 October 2019
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Applications of BBa_K2973009
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:
PCR reaction:
User Reviews
UNIQc242ec21b6d24c91-partinfo-00000006-QINU UNIQc242ec21b6d24c91-partinfo-00000007-QINU