Tissue microdissection and degenerate oligonucleotide primed-polymerase chain reaction (DOP-PCR) is an effective method to analyze genetic aberrations in invasive tumors.

Tissue microdissection and degenerate oligonucleotide primed-polymerase chain reaction (DOP-PCR) is an effective method to analyze genetic aberrations in invasive tumors.
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组织显微切割和简并寡核苷酸引物聚合酶链反应(DOP-PCR)是分析侵袭性肿瘤遗传畸变的有效方法。

DOI:
10.1016/s1525-1578(10)60653-8
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发表时间:
2001
期刊:
The Journal of molecular diagnostics : JMD
影响因子:
--
通讯作者:
Feuerstein,BG
Feuerstein,BG
中科院分区:
--
文献类型:
--
作者:
Hirose,Y;Aldape,K;Takahashi,M;Berger,MS;Feuerstein,BG

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我们扩增了来自冷冻 SF210 和 U251NCI 人胶质母细胞瘤细胞的不同量的 DNA,使用简并寡核苷酸引物 PCR (DOP-PCR) 产品作为测试探针进行比较基因组杂交 (CGH),并将结果与​​使用标准切口平移制备的探针进行的分析进行比较。接下来,我们从福尔马林固定和石蜡包埋的 U251NCI 细胞的苏木精-伊红 (HE) 和甲基绿 (MG) 染色、显微解剖切片中提取 DNA,通过 DOP-PCR 对其进行扩增和标记,并将其进行 CGH。最后,我们在来自单个人类混合神经胶质瘤组织的多个样本中使用了相同的方法。 50 pg 至 250 ng DNA 的 DOP-PCR 产物在生成与标准方法相同的 CGH 谱方面同样有效。来自 MG 染色细胞显微切片的 DOP-PCR 产物是 CGH 的有效探针,但 HE 染色的样品并不理想。随着 HE 染色样品比例的增加,CGH 特征恶化。来自神经胶质瘤组织 MG 染色的石蜡切片的显微切割片段的 DOP-PCR 产品产生了与其组织学特征相一致的 CGH 图谱。使用来自显微切割石蜡块的 DOP-PCR 产品进行 CGH 可以准确研究侵袭性肿瘤的细胞遗传学特征以及肿瘤组织内的细胞遗传学异质性。
We amplified various amounts of DNA derived from frozen SF210 and U251NCI human glioblastoma cells, carried out comparative genomic hybridization (CGH) using degenerate oligonucleotide primed-PCR (DOP-PCR) products as test probes, and compared results to analyses performed with probes prepared by standard nick translation. Next we extracted DNA from hematoxylin-eosin (HE)- and methyl green (MG)-stained, microdissected sections of formalin-fixed and paraffin-embedded U251NCI cells, amplified and labeled it by DOP-PCR, and subjected it to CGH. Finally, we used the same methods in multiple samples from a single human mixed glioma tissue. DOP-PCR products from 50 pg to 250 ng of DNA were equally effective in generating the same CGH profiles as the standard method. DOP-PCR products from microdissected pieces of MG-stained cells were effective probes for CGH, but HE-stained samples were not desirable. As the proportion of HE-stained sample increased, CGH profiles deteriorated. DOP-PCR products from microdissected pieces of MG-stained paraffin sections of glioma tissue produced CGH profiles compatible with their histological features. CGH performed with DOP-PCR products from microdissected paraffin blocks allows for the accurate investigation of the cytogenetic characteristics from invasive tumors and of cytogenetic heterogeneity within neoplastic tissue.
从石蜡包埋、福尔马林固定组织的小区域中分离出的 DNA 进行通用扩增。
DOI: --
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