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To knock out the 5’ splice site of Blustr, we co-transfected mESCs as described above, using a single sgRNA pZB plasmid and 200 ng of ssDNA oligonucleotide donor for homologous recombination (Extended Data Fig. 8c). The oligo was ordered as an ultramer from Integrated DNA Technologies (Table S2). We genotyped these insertions through amplicon sequencing using an Illumina MiSeq (primers in Table S2).
Copyright and License information: ©2016
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