518 15 404 10 10 0 5.28x10"" 1.78x10"" 394 11 11 0 10 0 12 0 0(399) (357)14 3 3 8/2000

524 15 397 10 10 0 2.70xl0~3 4.67xl0~s 394 11 11 0 9 0 12 0 1 (305)15 2 2 9/2000

525 15 397 10 10 0 6.95xl0~3 2.31 X 10~s 394 11 11 0 9 0 12 0 1 (305)15 2 2 9/2000

526 15 446 13 13 0 2.53x10"" 5.04xl0~s 329 15 15 0 13 0 15 1 2 (357)14 3 6 9/2000

a In all cases the known dam was confirmed by the microsatellite data. The putative sire was always one of the males used in the natural mating or AI. Giant pandas SB 495 and SB 509 have one mismatch with the putative sire, and this is probably due to the presence of a null allele for locus Ame-m25 (see text). Exclusion data are shown for all male pandas at the breeding station and for only those sires deemed possible from the breeding records; b Next most likely sires shown in parenthesis; c ND, not determined, no DNA available for dam.

(SB 298) is further excluded at Ame-p.14, because his two alleles cannot be combined with the alleles of Yong Ba to generate the Tian Tian genotype. (This exclusion is labelled in Table 10.3, column 14 under 'Off-Sire-Dam mismatch') Similarly, Lo Lo is excluded as a sire at loci Ame-m27 and Ame-p.70 due to Off-Sire loci mismatches. Pan Pan matches as the sire and fits with the dam at all loci tested; he is implicated as Tian Tian's sire. We conclude therefore that the natural mating by Pan Pan (SB 308) to Yong Ba (SB 397) produced the offspring Tian Tian (SB 458).

Validation of the genotyping results with blinded samples

To validate parentage analysis, 17 cubs born at the China Conservation and Research Centre for Giant Pandas (Wolong) in 1999 and 2000 were tested in a blinded fashion. Buccal swabs were obtained from each animal and given a code number. The scientists performing the DNA isolation and parentage testing were not given any information concerning the animals' possible parents. All breeding age males and females at the Wolong facility were checked as possible parents, although DNA samples were not available for some dams. Correct maternal assignments were made for every cub when the dam's DNA was available. For cubs in which the mother's DNA was absent, all available and tested females were excluded as potential dams. Four of the cubs were correctly inferred to be females since they were heterozygous for microsatellite Ame-^19. Five cubs were correctly inferred to be males based on the fact that they did not express an X chromosome allele from their sire at locus Ame-^19. In the case of Long Shen's (SB 518) paternity, two full brothers (SB 394 and SB 399) were both possible sires, and in the case of Long Xin (SB 516), a father/son pair were found to be possible sires (SB 308/413). Inspection of the breeding records made it possible to exclude all but a single sire in both of these instances (see Table 10.3). In all 17 cases, the implicated paternity from the blind analysis was consistent with one of the named possible sires based upon natural mating and AI.

Addressing unknown paternities

Tables 10.3 and 10.4 list the maternity/paternity assignments plus the parameters of microsatellite genotype allele matching for 50 cubs born at the Chengdu and Wolong facilities. For each paternity queried, the

Table 10.4. Microsatellite confirmed parentage at the Chengdu and Wolong breeding facilities
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