CD34 expansion media: StemSpan SFEM II (Stemcell Tech., 09655) with 100ng/ml TPO, SCF, IL-6, FLT-3
RNase decontamination solution (RNaseZap, Thermo Fisher, AM9780)
Chemically synthesized sgRNA, with 2'-O-methyl 3’phosphorothioate modifications for optimal performance. (Synthego)
Nuclease-free 1X TE Buffer (Tris-EDTA, pH 8.0)
Nuclease-free water
P3 4D-Nucleofector Kit (Lonza, 20ul Nucleocuvette strips format, V4XP-3032 or V4SP-3096,100ul Cuvette format, V4XP-3012)
sNLS-SpCas9-sNLS nuclease (Aldevron, cat. no. 9212-0.25MG)
CD34 expansion media: StemSpan SFEM II (Stemcell Tech., 09655) with 100ng/ml TPO, SCF, IL-6, FLT-3
gDNA column purification kit (E.Z.N.A. MicroElute Genomic DNA Kit, Omega Bio-Tek, cat. no. D3096)
High-fidelity DNA polymerase (Phusion High-Fidelity DNA Polymerase, NEB, cat. no. M0530L); dNTPs; PCR-grade DMSO
PCR and sequencing primers appropriate for the genomic region being targeted
PCR column purification kit (Monarch PCR & DNA Clean-up Kit, New England BioLabs, T1030S)