Single Primer PCR Fingerprinting For Rabbits

In conventional DNA fingerprinting, hypervariable and repetitive sequences ( minisatellite or microsatellite DNA ) are detected with hybridization probes. As demonstrated here, these probes can be used as single primers in the polymerase chain reaction (PCR) to generate individual fingerprints. Several conventional DNA fingerprinting probes were used to prime the PCR, yielding distinctive, hypervariable rmiltifragment profiles for different strains of rabbits. PCR fingerprinting with the oligonucleotide primers (GTG)5, (GACA)4, M13 core sequence (5-GAGGGTGGCGGTTCT-3') , generated DNA polymorphisms with all 4 strains of rabbits. Single primer amplified reactions(SPAR) techniques were used to investigate the genetic structures of the Vc-1 Rex rabbit population Vc-HRex rabbit population . RDB population and QZL population. Four DNA pools of the populations and 32 individual DNA samples were used. Three primers were screened and all of them were polymorphic, the band numbers and the fragment length were different in each primer. The fragment length of products from 3 primers in different samples were between 250 and 2000 bp, and the band numbers from primer M13 was between 6 and 11, primer (GTG)s was between 4 and 10, primer (GACA)4 was between 9 and 12, which were distinctly and easy to be distinguished. The fragment shared (F) and the genetic distance (D) were calculated based on it by using Phylogenetic Computer Tools soft among 4 populations and 32 individuals. The genetic distance indices were between 0.097 and 0.508 in 4 populations and between 0.036 and 0.480 in 32 individuals. The genetic distance between Vc-1 Rex rabbit and QZL was the furthest, Vc- I Rex rabbit and Vc- II Rex rabbit was the nearest. Amongindividuals in the Vc-1 Rex rabbit,Vc- II Rex rabbit,RDB and QZL population, the genetic distance indices were 0.036 - 0.176, 0.055 - 0.200, 0.037 -0.176,and 0.061- 0.217, respectively. The polygenetic trees were drawn, which showed the genetic relationships among 4 populations and 32 individuals objectively and explicitly.Amplification of polymorphic DNA patterns by PCR with these primers offer several advantages over classical DNA fingerprinting techniques, appear to be more reliable than PCR-based methods for detecting polymorphic DNA, such as analysis of random-amplified polymorphic DNA, and should be applicable to many other organisms.