Identification of TG mice and assessment of the integrity of BAC transgenes in TG mice.

Abstract

<p>(A) PCR for detecting TG mice. Two pairs of primers were used to amplify the 489 bp and 282 bp inserts. M, 100 bp DNA ladder; 10–37, TG founders; N, genomic DNA from WT mouse as a negative control; P, circular BAC DNA as a positive control. (B) Southern blot analysis of integrated pFSH transgenes. Purified genomic DNA (10 µg) from each TG founder was digested by <i>Bam</i>HI overnight at 37°C and analyzed by Southern blotting using probes specific for <i>pFSHα</i> and <i>pFSHβ</i>. 10–37, TG founders; P, porcine genomic DNA as positive control; N, genomic DNA from a WT mouse as a negative control. Band intensity was quantified by densitometry using Quantity-One software (Bio-Rad). (C–D) The two ends of both the BAC412H8 and BAC183O11 transgenes were amplified separately from TG mouse genomic DNA using 5′- or 3′-specific primers.</p