从干血斑中最大化脱氧核糖核酸产量。
Maximizing deoxyribonucleic acid yield from dried blood spots.
作者信息
Lane Julie A, Noble Janelle A
机构信息
Children's Hospital Oakland Research Institute, Oakland, California 94609 , USA.
出版信息
J Diabetes Sci Technol. 2010 Mar 1;4(2):250-4. doi: 10.1177/193229681000400204.
BACKGROUND
One source of deoxyribonucleic acid (DNA) for genetic studies is the utilization of dried blood spots stored on paper cards (Guthrie cards) collected shortly after birth. These cards represent an important source of material for epidemiologic and population-based genetic studies. Extraction of DNA from these cards can lead to variable amounts of recovered DNA. We report here results of our efforts to maximize yield from this valuable, but nonrenewable, resource.
METHOD
Commercial methods of DNA extraction from blood cards were used, and protocol modifications were introduced that enhanced DNA yield.
RESULTS
Use of a commercial solvent prior to DNA extraction steps gave greater yields than extraction without the solvent. Modification of the elution step by use of prewarmed extraction buffer and a soaking step at an elevated temperature increased yield by 6- to 10-fold.
CONCLUSIONS
The modified DNA extraction method yielded as much as 660 ng of DNA from a single 5-mm-diameter punch of a blood spot card. The DNA performed well in downstream, polymerase chain reaction-based applications.
背景
用于基因研究的脱氧核糖核酸（DNA）的一个来源是利用出生后不久采集并存储在纸卡（格思里卡片）上的干血斑。这些卡片是流行病学和基于人群的基因研究的重要材料来源。从这些卡片中提取DNA可能会导致回收的DNA量有所不同。我们在此报告我们为从这种宝贵但不可再生的资源中最大化产量所做努力的结果。
方法
采用从血卡中提取DNA的商业方法，并引入了可提高DNA产量的方案修改。
结果
在DNA提取步骤之前使用商业溶剂比不使用溶剂进行提取的产量更高。通过使用预热的提取缓冲液和在升高温度下的浸泡步骤对洗脱步骤进行修改，使产量提高了6至10倍。
结论
改进后的DNA提取方法从单个直径5毫米的血斑卡打孔中可获得多达660纳克的DNA。该DNA在基于聚合酶链反应的下游应用中表现良好。
Zotero 插件