Knock In Donor 1 Singlet
DNA was purified using GeneJet (K0721  ThermoFisher - Waltham, MA) according to the manufacturer's instructions. DNA purity and concentration was verified using JetSeq Library Quantification Lo-Rox kit (BIO 68029  Meridian BioScience  Memphis, TN) and Quant-iT PicoGreen (p7589  Invitrogen  Carlsbad, CA).  The NGS library was prepared using rhAmpSeq CRISPR Library Kit (cat # 10007318 - Integrated DNA Technologies - Coralville, Iowa). All samples were treated in each protocol as recommended by the manufacturer.  Briefly, 50ng of genomic DNA was added to the Targeted rhAmp PCR 1 which contains rhAmpSeq CRISPR forward and Reverse pools.  The post PCR1 product was 1:20 dilute and added to Indexing PCR 2 using Illumina Nextera XT primers (Illumina - San Diego, California).  Finally, libraries were pooled and purified with AMPure XP beads (Beckman Coulter - Indianapolis, IN) protocol.  Sequencing was performed using Illumina MiSeq 2X300 paired-end read length (Illumina - San Diego, California).
SUB15152635
KI_D1_Sing