Part:BBa_K4778005

ssDNA-F_D

Biology

Sequence：5‘-AAA AAA AAA ATAGT/i6FAMdT/CTTAT/iBHQ1dT/TCAG AAA AAA AAA-3’

Length: 32nt

It is the substrate of Cas12a_sgRNA_D_assistantDNA_D with ssDNA trans-cleaving activity in the 2023 project of AFMU_China team. When the structure of ssDNA-F_D remains normal, the distance between FAM and BHQ groups modified on it is 7nt, and the suppressor group can well inhibit the luminescence ability of fluorescent groups. When ssDNA-F was cleaved by Cas12a and broken, the distance between FAM and BHQ groups increased and lost its inhibitory effect. We use 492nm and 518nm as the excitation and reception wavelengths of the FAM group respectively. The fluorescence activity is activated and released, and the data are recorded and read out by the enzyme labeling instrument as the indication of nucleic acid signals.

Results

Fig1，Detection of different concentrations of miRNA in 50uL reaction system When ssDNA-F is not cleaved, it has a good negative control effect, and when it is cleaved, it can release obvious fluorescence quickly. Its characteristics enable it to bear the role of signal display.

Fig2,The LC-MS data of ssDNA-F_D.