Walker Jerilyn A, Hughes David A, Hedges Dale J, Anders Bridget A, Laborde Meredith E, Shewale Jaiprakash, Sinha Sudhir K, Batzer Mark A
Department of Biological Sciences, Biological Computation and Visualization Center, Louisiana State University, 202 Life Sciences Building, Baton Rouge, LA 70803, USA.
Genomics. 2004 Mar;83(3):518-27. doi: 10.1016/j.ygeno.2003.09.003.
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.
我们设计并评估了一系列基于聚合酶链反应（PCR）的检测方法，这些方法针对特定类别（鸟类）、特定目（啮齿目）以及特定物种（马、犬、猫、大鼠、仓鼠、豚鼠和兔），利用基因组特异性短散在元件和长散在元件的扩增来鉴定和定量DNA。使用基于SYBR Green的检测方法，这些检测的最低有效定量水平在起始DNA模板0.1 ng至0.1 pg之间。在PCR进行30个循环之前，来自其他16个物种的DNA模板的背景交叉扩增可忽略不计。PCR扩增子的物种特异性通过检测方法从混合（复杂）来源中准确检测已知量的物种特异性DNA的能力得到进一步证明。本文报道的10种检测方法将有助于从复杂生物材料中灵敏地检测和定量常见家畜和鸟类物种的DNA。
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