Nucleospin tissue genomic dna extraction kit

Genomic DNA from pools of >10 individuals of sds3-Cas9 and all shu-Cas9 isolines was extracted using the NucleoSpin Tissue genomic DNA extraction kit (Machery-Nagel). Genomic DNA was digested with either BamHI-HF (NEB R3136), MspI (NEB R0106), or NcoI-HF (NEB R3193). Adapters (Supplementary Table S6) were ligated with T4 DNA ligase (NEB M0202) overnight at 14 °C. Primary and nested PCRs were carried out using DreamTaq polymerase (Thermo Fisher Scientific EP0712) using the primers listed in Supplementary Table S6. Amplicons were purified using NucleoSpin Gel and PCR Clean-up Kit and Sanger sequenced using the primers listed in Supplementary Table S6. Genomic locations are listed in Supplementary Table S5.

Genomic DNAs were isolated and purified from leukocytes using the NucleoSpin Tissue genomic DNA extraction kit (Machery-Nagel, Germany). DNA samples were processed by the Illumina Infinium method using the HumanCytoSNP—12 v2.1 (Illumina, San Diego, CA, USA) microarray platform and analysed using KaryoStudio v1.4 software (Illumina). Confirmation of the null deletions and cascade testing of the parents was performed using Affymetrix CytoScan 750K array using the manufacture’s protocols and analysed using Chromosome Analysis Suite vCytoB-N1.2.2.271 (Affymetrix, Thermo Fisher Scientific).