Quantitative PCR for DNA identification based on genome-specific interspersed repetitive elements.
Quantitative PCR for DNA identification based on genome-specific interspersed repetitive elements.
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DOI：
10.1016/j.ygeno.2003.09.003
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发表时间：
2004-03
期刊：
影响因子：
4.4
通讯作者：
Jerilyn A. Walker;David A Hughes;D. Hedges;Bridget A Anders;Meredith E Laborde;J. Shewale;S. Sinha;M. Batzer
中科院分区：
文献类型：
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作者：
Jerilyn A. Walker;David A Hughes;D. Hedges;Bridget A Anders;Meredith E Laborde;J. Shewale;S. Sinha;M. Batzer
我们设计并评估了一系列基于类特异性 (Aves)、目特异性 (Rodentia) 和物种特异性（马、犬、猫、大鼠、仓鼠、豚鼠和兔）聚合酶链式反应 (PCR) 的检测方法，通过扩增基因组特异性短和长散布元件来鉴定和定量 DNA。使用基于 SYBR Green 的检测，检测的最低有效定量水平范围为 0.1 ng 至 0.1 pg 起始 DNA 模板。在 30 个 PCR 循环之前，与来自其他 16 个物种的 DNA 模板的背景交叉扩增可以忽略不计。 PCR 扩增子的物种特异性通过该检测准确检测来自混合（复杂）来源的已知数量的物种特异性 DNA 的能力得到进一步证明。这里报道的 10 种检测将有助于促进复杂生物材料中常见家畜和鸟类 DNA 的灵敏检测和定量。
We have designed and evaluated a series of class-specific (Aves), order-specific (Rodentia), and species-specific (equine, canine, feline, rat, hamster, guinea pig, and rabbit) polymerase chain reaction (PCR)-based assays for the identification and quantitation of DNA using amplification of genome-specific short and long interspersed elements. Using SYBR Green-based detection, the minimum effective quantitation levels of the assays ranged from 0.1 ng to 0.1 pg of starting DNA template. Background cross-amplification with DNA templates derived from sixteen other species was negligible prior to 30 cycles of PCR. The species-specificity of the PCR amplicons was further demonstrated by the ability of the assays to accurately detect known quantities of species-specific DNA from mixed (complex) sources. The 10 assays reported here will help facilitate the sensitive detection and quantitation of common domestic animal and bird species DNA from complex biomaterials.