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. 2022 Oct 31;10:e14283. doi: 10.7717/peerj.14283

Figure 4. Experimental verification of primers selected by the script.

Figure 4

Lane M, DNA marker; lane 1, PCR amplification of ECdnaQ with primers gene_5/overlap_3 from pETDuet-1-ECdnaQ; lane 2, PCR amplification of TBdnaN with primers overlap_5 and gene_3 from pET28a-TBdnaN; lane 3, gel recycling of the ECdnaQ band from lane 2; lane 4, gel recycling of the TBdnaN band from lane3; lane 5, PCR amplification of full-length fragment with primers gene_5/gene_3 from mixed template DNA of ECdnaQ and TBdnaN; lane 6, producing full-length fragment via overlap extension without primers gene_5/gene_3; lane 7, negative control, the same as lane 5 but using ddw as template. The lowest arrow in the right side indicates the position of ECdnaQ. The middle arrow in the right side indicates the position of TBdnaN. The highest arrow in the right side indicates the position of full-length chimera fragment.