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. 2012 Nov 28;7(11):e49450. doi: 10.1371/journal.pone.0049450

Figure 3. Analysis of the F8 in cloned piglets.

Figure 3

(A) PCR analysis of genomic DNA of piglet DNA was shown. Genomic DNA of wild-type, 134-fetus, piglet #1, piglet #2, piglet #3, and piglet #4 was subjected to PCR analysis with primers Exon 14 sF and Exon 18 sR as in Figure 1. The 8.3 kb exon 14–18 band was amplified from the 134-fetus DNA and the cloned piglet DNA. (B) Southern blotting with a 5′ exon 14 probe (on Sac I− or Sac I + Stu I-digested DNA) showed the same mobility shifts of the bands as those in Figure 2B and confirmed the insertion of the Neo resistant gene in F8 of the cloned piglets. (C) RT-PCR analysis of piglet liver RNA was shown. Two independent PCRs (exons 14–16 and exons 18–22) revealed the absence of FVIII mRNA from the liver of cloned piglet #3. Control GAPDH mRNA was detected in the liver RNA of piglet #3 as in the wild type (WT).