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For the KO genomic DNA validation, the QuickExtract™ DNA Extraction Solution 1.0 kit was used. Briefly, cells were harvested, extraction solution was added and tubes were vortexed for 15 s. Then, incubated for 6 min at 65 °C, vortexed again and incubated for 2 min at 98 °C. Five microliters was taken for PCR. We used the following primers, KO#1 (gRNA #1) Fw 5′-TTAGGCGCGCCAAGAGTTTTGGGGGGATAGTTT-3′ Rev 5′-GCCTTAATTAAAAAATTGTATCATTTTAGCACCC-3′ and KO#2 (gRNA #2) Fw 5′-TTAGGCGCGCCTTAAGGCAAAGTGGGGAGAGAATA-3′ Rev 5′-GCCTTAATTAAGGAAGCTCACAATTACTTCACCC-3′.
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