Assessment of DNA extracted from FTA cards for use on the Illumina iSelect BeadChip.
Assessment of DNA extracted from FTA cards for use on the Illumina iSelect BeadChip.
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DOI：
10.1186/1756-0500-2-107
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发表时间：
2009-06-16
影响因子：
1.8
通讯作者：
Taylor, Jeremy F
中科院分区：
文献类型：
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作者：
McClure, Matthew C;McKay, Stephanie D;Taylor, Jeremy F
背景技术背景：由于FTA卡为DNA的现场收集提供了理想的介质，我们试图评估从该来源提取的基因组DNA的质量，以用于Illumina BovineSNP 50 iSelect BeadChip，其需要未结合的、相对完整的（片段大小>或= 2kb）和高质量的DNA。使用市售GenSolve试剂盒（稍作修改）提取FTA卡上采集的牛血和鼻拭子样本。结果：方差分析表明，从FTA卡中提取的DNA与从PCI全血中提取的DNA之间的牛SNP50基因型检出率无显著差异（P > 0.72）。两个样本的t检验表明，从FTA卡中提取的DNA产生的基因型调用率和一致率与从全血中提取的DNA样本通过PCI检测产生的结果没有差异。我们得出结论，通过GenSolve试剂盒从FTA卡中提取的DNA具有足够高的质量，从而在Illumina iSelect分析时产生与从全血中提取的DNA获得的结果相当的结果技术.此外，我们验证了使用鼻拭子作为来自动物受试者的静脉血或口腔样本的替代品，以在该平台上可靠地产生高质量的基因型。
BACKGROUND: As FTA cards provide an ideal medium for the field collection of DNA we sought to assess the quality of genomic DNA extracted from this source for use on the Illumina BovineSNP50 iSelect BeadChip which requires unbound, relatively intact (fragment sizes >or= 2 kb), and high-quality DNA. Bovine blood and nasal swab samples collected on FTA cards were extracted using the commercially available GenSolve kit with a minor modification. The call rate and concordance of genotypes from each sample were compared to those obtained from whole blood samples extracted by standard PCI extraction.FINDINGS: An ANOVA analysis indicated no significant difference (P > 0.72) in BovineSNP50 genotype call rate between DNA extracted from FTA cards by the GenSolve kit or extracted from whole blood by PCI. Two sample t-tests demonstrated that the DNA extracted from the FTA cards produced genotype call and concordance rates that were not different to those produced by assaying DNA samples extracted by PCI from whole blood.CONCLUSION: We conclude that DNA extracted from FTA cards by the GenSolve kit is of sufficiently high quality to produce results comparable to those obtained from DNA extracted from whole blood when assayed by the Illumina iSelect technology. Additionally, we validate the use of nasal swabs as an alternative to venous blood or buccal samples from animal subjects for reliably producing high quality genotypes on this platform.