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This process requires two essential amino acids (lysine and methionine), iron (Fe 2+ ), vitamin B 6 in the form of pyridoxal 5′-phosphate, niacin in the form of nicotinamide adenine dinucleotide (NAD), and may also require vitamin C (ascorbate) (4)

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Per HDR reaction, 3.75 μl of 1 μM gRNA, 0.125 μl of 5 μg/μl TrueCut Cas9 v2 (A36498