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. 2021 Jul 14;37:33–41. doi: 10.1016/j.jare.2021.07.006

Fig. 3.

Fig. 3

Construction of gRNA expression Cassette. (A) Shows the restriction analysis of clones from the ligation of dsOligo at the BtgZ1 site of pENTR4-gRNA1 after digestion with BamH1 restriction enzyme. The clones without the 350 bp fragment are positive clones. (B) Shows the confirmation of insertion of second dsOligo at the Bsa1 site of pENTR4-gRNA1 after digestion with Bsa1 and BamH1 restriction enzymes. The positive clones are without digested fragment of 790 bp. (C) Shows the confirmation of Gateway LR recombination reaction after digestion of clones with HindIII restriction enzyme. The positive clones show the digestion pattern of 12 kb, 2.6 kb and 1 kb. (M, 1 kb DNA marker, C, control plasmid).