Difference between revisions of "Part:BBa K2973017"

Revision as of 13:02, 21 October 2019

IS6110 Reverse Primer with 5' Overhang

IS6110 is an insertion element found exclusively within the members of the Mycobacterium tuberculosis complex (MTBC). This part is a reverse primer with a trigger sequence on its 5' end. The primer is designed to amplify the IS6110 gene (BBa_K2973009), along with its forward primer (BBa_K2973016).

Usage and Biology

In our project, this primer (BBa_K2973017) was used along with its forward primer (BBa_K2973016) 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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Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2973017 short</partinfo>
-IS6110 is an insertion element found exclusively within the members of the Mycobacterium tuberculosis complex (MTBC). This part is a reverse primer with a trigger sequence on its 5' end. The primer is designed to amplify the IS6110 gene (BBa_K2973009), along with its forward primer (BBa_K2973016).
+IS6110 is an insertion element found exclusively within the members of the Mycobacterium tuberculosis complex (MTBC). This part is a reverse primer with a trigger sequence on its 5' end. The primer is designed to amplify the IS6110 gene (<partinfo>BBa_K2973009</partinfo>), along with its forward primer (<partinfo>BBa_K2973016</partinfo>).
 ===Usage and Biology===
-In our project, this primer (BBa_K2973017) was used along with its forward primer (BBa_K2973016) for the amplification reactions of the IS6110 gene ([[Part: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.
+In our project, this primer (<partinfo>BBa_K2973017</partinfo>) was used along with its forward primer (<partinfo>BBa_K2973016</partinfo>) for the amplification reactions of the IS6110 gene (<partinfo>BBa_K2973009</partinfo>) 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 (<partinfo>BBa_K2973007</partinfo>). The expected amplified product length is 136 bp.