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. 2013 Jul 29;2:138. Originally published 2013 Jun 10. [Version 2] doi: 10.12688/f1000research.2-138.v2

Figure 3. Locating the APHVIII flanking genomic sequence in 5A7.

Figure 3.

( A) A diagram showing a truncated pBC1 illustrating the APHVIII end of the linearized pBC1 vector. Primers used for PCR are shown by numbered black arrows. Thermal Asymmetric InterLaced 1 (TAIL1) PCR was performed using primer 4R and AD2. ( B) TAIL2 PCR was performed using primer 3R and AD2. Lanes 1 and 4 are zero DNA lanes; in lane 2, a 10-fold diluted TAIL1 PCR product was used for TAIL2 PCR; in lane 5, a 25-fold diluted TAIL1 PCR product was used for TAIL2 PCR; lanes 3 and 6 are blank lanes. The 850 bp product used for DNA sequencing is highlighted. ( C) Gel purified DNA product (850 bp) from the TAIL2 PCR was used to verify if the product was specific to the APHVIII gene. F and R stand for forward and reverse primers, respectively. AD2 is a non-degenerate primer. PCR primer names are labeled on the top of the gel. In lanes A and B, where triple primers were used for PCR, PCR products are labeled by the corresponding primer combinations that gave rise to the specific product. PCR product sizes are shown beside the primer combinations. All primer sequences are shown in Table 1. ST stands for 1 kb plus ladder (Invitrogen, Carlsbad, CA). DNA samples were run on a 1% agarose gel.