Chunk #78 — PROCEDURE — Functional testing: detection of genomic microdeletions by PCR ● TIMING 3–4 h hands-on; 2–3 weeks expansion

Source

Genome engineering using the CRISPR-Cas9 system.

Embedded

yes

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As with SURVEYOR assays, include a negative (untransfected sample) control. Set up the PCR as follows: ComponentAmount (μl)Final concentrationHerculase II PCR buffer, 5×101×dNTP, 100 mM (25 mM each)12 mMOut-Fwd primer, 10 μM10.2 μMOut-Rev primer, 10 μM10.2 μMHerculase II fusion polymerase1MgCl2, 25 mM21 mMDNA template12 ng μl−1ddH2O33Total50