The second stage of assessing the risk of horizontal transmission of the CRISPR construct will be in vitro, i.e. through assays that replicate the CRISPR/Cas9 system without the use of living organisms, but which provide experimental data for assessing the specificity of the system. This process will involve the application of techniques such as DNA extraction, PCR, enzymatic reactions, electrophoresis, densitometry, among others.
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