Khokhar Shama Khan, Mitui Midori, Leos Nora Kristine, Rogers Beverly Barton, Park Jason Y
Department of Pathology, Children's Medical Center, Dallas, TX, USA.
Clin Chem Lab Med. 2011 Oct 25;50(2):267-72. doi: 10.1515/CCLM.2011.763.
The aim of the study was to assess the performance of Promega, Maxwell® 16 for the extraction of genomic DNA from whole blood and FFPE tissue.
DNA was extracted from 10 whole blood and 10 FFPE specimens using six different commercial kits.
For whole blood, the mean DNA concentration obtained by Maxwell® 16 was significantly greater than either easyMAG® (p<0.0001) or QIAamp® Blood DNA kit (p<0.001). For FFPE, the mean DNA concentration obtained by the AllPrep® FFPE specific DNA/RNA kit was significantly greater than either the Maxwell® 16 (p<0.0001) or the general AllPrep® DNA/RNA kit (p<0.0001).
Comparative evaluation of the six DNA extraction kits indicated that the semi-automated Maxwell® 16 was superior for whole blood extraction while the manual AllPrep® FFPE DNA/RNA kit (Qiagen) performed better for FFPE DNA extraction in terms of quantity of DNA obtained. All six extraction methods (blood and FFPE) performed well in terms of purity. Although there were variances in the quantity of DNA obtained, there were no significant differences in the efficiency of these methods in yielding amplifiable DNA extracts, as demonstrated by β-actin for whole blood specimens. In evaluation of FFPE DNA extraction methods, the Qiagen AllPrep® FFPE DNA/RNA Mini Kit was the best for applications requiring larger amplicons, but for smaller amplicons the Maxwell was most consistent.
本研究的目的是评估普洛麦格公司的Maxwell® 16从全血和福尔马林固定石蜡包埋(FFPE)组织中提取基因组DNA的性能。
使用六种不同的商业试剂盒从10份全血和10份FFPE标本中提取DNA。
对于全血,Maxwell® 16获得的平均DNA浓度显著高于easyMAG®(p<0.0001)或QIAamp®血液DNA试剂盒(p<0.001)。对于FFPE,AllPrep® FFPE特异性DNA/RNA试剂盒获得的平均DNA浓度显著高于Maxwell® 16(p<0.0001)或通用的AllPrep® DNA/RNA试剂盒(p<0.0001)。
六种DNA提取试剂盒的比较评估表明,半自动的Maxwell® 16在全血提取方面更具优势,而手动的AllPrep® FFPE DNA/RNA试剂盒(Qiagen)在FFPE DNA提取方面,就获得的DNA量而言表现更好。所有六种提取方法(血液和FFPE)在纯度方面表现良好。尽管获得的DNA量存在差异,但如全血标本的β-肌动蛋白所示,这些方法在产生可扩增DNA提取物的效率上没有显著差异。在评估FFPE DNA提取方法时,Qiagen AllPrep® FFPE DNA/RNA Mini试剂盒最适合需要更大扩增子的应用,但对于较小的扩增子,Maxwell最为稳定。