Part:BBa_K4778007
probe-α
Usage and Biology
Sequence：5'-AGACACTGCCAUAAUUUCUACUAAGUGUAGAUGATCGTTACGCTAACACAACCAGCTA-3'
Length：58nt
Fig1.probe-α
This component is the first generation of miR-34a probe we designed. The yellow sequence is deoxyribonucleotides with a length of 22nt, and the blue sequence is ribonucleotides with a length of 36nt. In the first step of the reaction, the DNA sequence of the probe is complementary to the miR to achieve the identification of miR (Fig2). In the second-generation system, we also replaced scgRNA-F in the second reaction with probe-α. When the trans cleavage activity of Cas12a is activated, the single-stranded DNA of the probe can be cleaved, releasing new sgRNA, thereby activating new Cas12a, achieving exponential amplification of the signal (Fig3).
Fig2.The first generation system.
Fig3.The second generation system.
Results
The urea denaturing PAGE showed that we used the probe and DSN enzyme to recognize miR-34a-5p and release sgRNA. This shows that our probe is effective.（Fig4）
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