Fig. 3From: Customized one-step preparation of sgRNA transcription templates via overlapping PCR Using short primers and its application in vitro and in vivo gene editinga The electropherogram of DNA fragment amplified by the four overlapped primers. M represents 50 bp DNA marker. NC represents the negative control. (1) the tracr-R primer; (2) the two primers : tracr-R and AF3; (3) the three primers: tracr-R, AF3 and AF2; (4, 5, 6) the templates of EGFR, pu57-1 and pu57-2 amplified by four primers respectively. b–d represent the sequencing results of the transcription template of EGFR-Exon19, pu57-1, pu57-2 respectively. The underlined part represents the designed sequence of sgRNABack to article page