Part:BBa_K2973016

IS6110 Forward Primer with 5' Overhang

IS6110 is an insertion element found exclusively within the members of the Mycobacterium tuberculosi complex (MTBC). This part is a forward primer with a T7 Promoter (BBa_J64997) on its 5' end. The primer is designed to amplify the IS6110 gene (BBa_K2973009), along with its reverse primer (BBa_K2973017).

Usage and Biology

In our project, this primer (BBa_K2973016) was used along with its reverse primer (BBa_K2973017) 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.

RPA reaction after clean up of the amplified product:

PCR reaction:

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]