Mitsouras Katherine, Faulhaber Erica A
College of Osteopathic Medicine of the Pacific, Western University of Health Sciences, Pomona, CA, USA.
BMC Res Notes. 2009 Oct 29;2:219. doi: 10.1186/1756-0500-2-219.
The domestic dog presents an attractive model system for the study of the genetic basis of disease. The development of resources such as the canine genome sequence and SNP genotyping platforms has allowed for the implementation of canine genetic studies. Successful implementation of such studies depends not only on the quality of individual DNA samples, but also on the number of samples obtained. The latter can be maximized using a non-invasive DNA collection method that can increase study participation. We compared the DNA yield and quality obtained from blood and buccal swabs to those obtained using a non-invasive saliva collection kit (Oragene *ANIMAL kit). We also assessed the success rate of PCR amplification and genotyping accuracy of DNA isolated using these collection methods.
Comparison of DNA yields from matched saliva, blood and buccal swab samples showed that yields from saliva were significantly higher than those from blood (p = 0.0198) or buccal swabs (p = 0.0008). Electrophoretic analysis revealed that blood and saliva produced higher quality DNA than buccal swabs. In addition, a 1.1-kb PCR fragment was successfully amplified using the paired DNA samples and genotyping by PCR-RFLP yielded identical results.
We demonstrate that DNA yields from canine saliva are higher than those from blood or buccal swabs. The quality of DNA extracted from saliva is sufficient for successful amplification of a 1.1-kb fragment and for accurate SNP genotyping by PCR-RFLP. We conclude that saliva presents a non-invasive alternative source of high quantities of canine genomic DNA suitable for genotyping studies.
家犬是研究疾病遗传基础的一个有吸引力的模型系统。犬类基因组序列和单核苷酸多态性(SNP)基因分型平台等资源的开发使得犬类遗传学研究得以开展。此类研究的成功实施不仅取决于单个DNA样本的质量,还取决于所获得样本的数量。使用一种可增加研究参与度的非侵入性DNA采集方法可使样本数量最大化。我们比较了从血液和口腔拭子中获得的DNA产量和质量与使用非侵入性唾液采集试剂盒(Oragene *ANIMAL试剂盒)所获得的DNA产量和质量。我们还评估了使用这些采集方法分离的DNA的PCR扩增成功率和基因分型准确性。
对匹配的唾液、血液和口腔拭子样本的DNA产量进行比较,结果显示唾液中的产量显著高于血液中的产量(p = 0.0198)或口腔拭子中的产量(p = 0.0008)。电泳分析表明,血液和唾液产生的DNA质量高于口腔拭子。此外,使用配对的DNA样本成功扩增出了一个1.1 kb的PCR片段,并且通过PCR-RFLP进行基因分型得到了相同的结果。
我们证明犬类唾液中的DNA产量高于血液或口腔拭子中的产量。从唾液中提取的DNA质量足以成功扩增一个1.1 kb的片段,并通过PCR-RFLP进行准确的SNP基因分型。我们得出结论,唾液是一种非侵入性的替代来源,可提供大量适合基因分型研究的犬类基因组DNA。