• 主题
高级搜索  >
历史搜索 清空历史
首页> 美国卫生研究院文献>ISRN Oncology >Comparison of Whole Genome Amplification Methods for Analysis of DNA Extracted from Microdissected Early Breast Lesions in Formalin-Fixed Paraffin-Embedded Tissue
【2h】

Comparison of Whole Genome Amplification Methods for Analysis of DNA Extracted from Microdissected Early Breast Lesions in Formalin-Fixed Paraffin-Embedded Tissue

机译：全基因组扩增方法分析从福尔马林固定石蜡包埋组织中的显微解剖早期乳腺病变中提取的DNA的比较

代理获取
本网站仅为用户提供外文OA文献查询和代理获取服务，本网站没有原文。下单后我们将采用程序或人工为您竭诚获取高质量的原文，但由于OA文献来源多样且变更频繁，仍可能出现获取不到、文献不完整或与标题不符等情况，如果获取不到我们将提供退款服务。请知悉。
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

To understand cancer progression, it is desirable to study the earliest stages of its development, which are often microscopic lesions. Array comparative genomic hybridization (aCGH) is a valuable high-throughput molecular approach for discovering DNA copy number changes; however, it requires a relatively large amount of DNA, which is difficult to obtain from microdissected lesions. Whole genome amplification (WGA) methods were developed to increase DNA quantity; however their reproducibility, fidelity, and suitability for formalin-fixed paraffin-embedded (FFPE) samples are questioned. Using aCGH analysis, we compared two widely used approaches for WGA: single cell comparative genomic hybridization protocol (SCOMP) and degenerate oligonucleotide primed PCR (DOP-PCR). Cancer cell line and microdissected FFPE breast cancer DNA samples were amplified by the two WGA methods and subjected to aCGH. The genomic profiles of amplified DNA were compared with those of non-amplified controls by four analytic methods and validated by quantitative PCR (Q-PCR). We found that SCOMP-amplified samples had close similarity to non-amplified controls with concordance rates close to those of reference tests, while DOP-amplified samples had a statistically significant amount of changes. SCOMP is able to amplify small amounts of DNA extracted from FFPE samples and provides quality of aCGH data similar to non-amplified samples.
机译：为了了解癌症的进展，希望研究其发展的最早阶段，通常是微观病变。阵列比较基因组杂交（aCGH）是发现DNA拷贝数变化的有价值的高通量分子方法。然而，它需要相对大量的DNA，这很难从显微切割的病变中获得。开发了全基因组扩增（WGA）方法以增加DNA数量；但是，它们对福尔马林固定石蜡包埋（FFPE）样品的重现性，保真度和适用性提出了质疑。使用aCGH分析，我们比较了WGA的两种广泛使用的方法：单细胞比较基因组杂交方案（SCOMP）和简并寡核苷酸引发的PCR（DOP-PCR）。通过两种WGA方法扩增癌细胞系和显微解剖的FFPE乳腺癌DNA样品，并进行aCGH。通过四种分析方法将扩增的DNA的基因组图谱与未扩增的对照的基因组图谱进行比较，并通过定量PCR（Q-PCR）进行验证。我们发现，SCOMP扩增的样品与未扩增的对照非常相似，符合率接近参考测试，而DOP扩增的样品具有统计学上的显着变化。 SCOMP能够扩增从FFPE样品中提取的少量DNA，并提供与未扩增样品相似的aCGH数据质量。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
代理获取

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号