Multiplex loss of heterozygosity analysis by using single or very few cells.
Multiplex loss of heterozygosity analysis by using single or very few cells.
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使用单个或很少的细胞进行多重杂合性丢失分析。
DOI：
10.1016/s1525-1578(10)60699-x
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
Li,Honghua
中科院分区：
文献类型：
--
作者：
Cui,Xiangfeng;Feiner,Helen;Li,Honghua
杂合性丢失（LOH）是抑癌基因失活的标志。然而，杂合性丢失分析仅限于小规模或很少的遗传标记。为了显着扩大研究规模并在分析中纳入大量标记，建立了使用单细胞或单细胞等效物同时输入 10 个标记的实验条件。在此条件下，使用单个组织培养的人类细胞时，等位基因扩增失败率为3.7%。当使用来自 5 μm 石蜡存档的乳腺肿瘤组织切片的 30 个细胞时，五个杂合位点中的四个的失败率为 0%，第五个为 10%。当使用来自石蜡存档的乳腺组织的 5 或 10 个细胞时，观察到较小的扩增失败率（平均 6.1% 和 6.7%）。这些结果表明，利用高质量的聚合酶链反应 (PCR) 引物，使用新鲜组织中的单个细胞或石蜡存档标本中的极少数细胞可以获得可靠的结果。结果还表明，当使用有限数量的材料进行测定时，包括重复、使用高质量引物以及优化 PCR 条件的重要性。证明了用很少的石蜡包埋乳腺癌组织进行 LOH 分析的可行性。
Loss of heterozygosity (LOH) is an indication of tumor suppressor gene inactivation. However, loss of heterozygosity analysis has been limited to either a small scale or to very few genetic markers. To significantly increase the scale of study and to include a large number of markers in the analysis, experimental conditions were established for using single cells or single cell equivalent with 10 markers typed simultaneously. Under these conditions, the allele amplification failure rate was 3.7% when single tissue cultured human cells were used. When 30 cells from a 5-μm paraffin-archived breast tumor tissue section were used, the failure rates were 0% for four of the five heterozygous loci and 10% for the fifth. Small amplification failure rates (6.1% and 6.7% on average) were observed when 5 or 10 cells from paraffin-archived breast tissue were used. These results indicate that with polymerase chain reaction (PCR) primers of high quality, it is possible to obtain reliable results by using single cells from fresh tissue or very few cells from paraffin-archived specimens. The results also show the importance of including replicates, using primers of high quality, and optimizing PCR conditions when a limited amount of material is used for the assay. The feasibility of LOH analysis with very few paraffin-embedded breast cancer tissues was demonstrated.
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人免疫球蛋白重链区 DNA 序列多态性簇的多重基因型测定。
影响因子：
4.4
作者：
H. Li;L. Hood
通讯作者：
L. Hood
DOI：
10.1073/pnas.89.13.5847
发表时间：
1992-07-01
影响因子：
11.1
作者：
ZHANG, L;CUI, XF;ARNHEIM, N
通讯作者：
ARNHEIM, N
影响因子：
2.5
作者：
Larsen, M;Ressler, SJ;Rowley, DR
通讯作者：
Rowley, DR
DOI：
10.1073/pnas.87.12.4580
发表时间：
1990-06-01
影响因子：
11.1
作者：
LI, HH;CUI, XF;ARNHEIM, N
通讯作者：
ARNHEIM, N
DOI：
10.1159/000133886
发表时间：
1995-01-01
期刊：
CYTOGENETICS AND CELL GENETICS
影响因子：
--
作者：
CLETONJANSEN, AM;MOERLAND, HW;CORNELISSE, CJ
通讯作者：
CORNELISSE, CJ