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| 通过石蜡包埋组织标本构建肝细胞癌基因组文库的研究 | |
| 引用本文： | 肖建,于倩,范露丹,姚利民,卢川,马韵,曾丽霞,李运千,黄永秩,黄小英,黄炳臣,姚金光,黄照权,龙喜带. 通过石蜡包埋组织标本构建肝细胞癌基因组文库的研究[J]. 右江民族医学院学报, 2012, 34(1): 4-6. DOI: 10.3969/j.issn.1001-5817.2012.01.002 |
| 作者姓名： | 肖建 于倩 范露丹 姚利民 卢川 马韵 曾丽霞 李运千 黄永秩 黄小英 黄炳臣 姚金光 黄照权 龙喜带 |
| 作者单位： | 1. 右江民族医学院2009级临床医学专业,广西百色,533000 2. 右江民族医学院2008级医学影像学专业,广西百色,533000 3. 广西医科大学附属肿瘤医院病理科,广西南宁,530027 4. 广西医科大学附属医院病理科,广西南宁,530023 5. 桂林医学院附属医院病理科,广西桂林,541000 6. 右江民族医学院附属医院,广西百色,533000 |
| 基金项目： | 国家自然科学基金资助项目，广西自然科学基金资助项目，广西高校重点建设实验室专项课题 |
| 摘 要： | 目的探讨通过石蜡包埋组织标本构建肝细胞癌(hepatocellular carcinoma,HCC)肿瘤组织基因组DNA文库的方法。方法收集符合要求的石蜡包埋HCC肿瘤组织标本,运用组织裂解改良法提取基因组DNA,构建HCC肿瘤组织基因组DNA文库,通过SmartSpecTM核酸蛋白测量法、TaqMan-MGB-PCR及琼脂糖凝胶电泳法等分析评估所得文库质量。结果本研究共收集到2 108例合格石蜡包埋HCC肿瘤组织标本,构建了HCC组织标本基因组DNA文库,经质量监控分析所得DNA的浓度大于或接近0.1μg/μl,对应A260/A280比值介于1.6~2.0,TaqMan-MGB-PCR实时定量曲线光滑、波峰单一,琼脂糖凝胶电泳条带单一、清晰、无拖尾现象,所构建的文库DNA质量较好,与对照DNA质量相比差异无显著性(P>0.05)。结论通过石蜡包埋组织标本可构建用于HCC研究的肿瘤组织基因组DNA文库。 |
| 关 键 词： | 石蜡包埋 癌,肝细胞 基因组DNA文库 质量控制 |
The construction of genomic DNA library with hepatocellular carcinoma neoplasm tissues embedded by paraffin |
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| XIAO Jian,YU Qian,FAN Lu-dan,YAO Li-min,LU Chuan,MA Yun,ZENG Li-xia,LI Yun-qian,HUANG Yong-zhi,HUANG Xiao-ying,HUANG Bing-chen,YAO Jin-guang,Huang Zhao-quan,LONG Xi-dai. The construction of genomic DNA library with hepatocellular carcinoma neoplasm tissues embedded by paraffin[J]. Journal of Youjiang Medical College For Nationalities, 2012, 34(1): 4-6. DOI: 10.3969/j.issn.1001-5817.2012.01.002 | |
| Authors: | XIAO Jian YU Qian FAN Lu-dan YAO Li-min LU Chuan MA Yun ZENG Li-xia LI Yun-qian HUANG Yong-zhi HUANG Xiao-ying HUANG Bing-chen YAO Jin-guang Huang Zhao-quan LONG Xi-dai |
| Affiliation: | 1.Grade 2009,Department of Clinical Medicine,Youjiang Medical University for Nationalities,Baise,Guangxi 533000,China 2.Grade 2008,Department of Imaging Medicine,Youjiang Medical University for Nationalities,Baise,Guangxi 533000,China;3.Department of Pathology,the Affiliated Tumor Hospital of Guangxi Medical University,Nanning,Guangxi 530027,China;4.Department of Pathology,the Affiliated Hospital of Guangxi Medical University,Nanning,Guangxi 530023,China;5.Department of Pathology,the Affiliated Hospital of Guilin Medical College,Guilin,Guangxi 541000,China;6.Department of Pathology,Affiliated Hospital of Youjiang Medical University forNationalities,Baise,Guangxi 533000,China) |
| Abstract: | Objective To develop a procedure for construction of genomic DNA library with paraffin-embedded neoplastic tissue specimens from patients with hepatocellular carcinoma(HCC).Methods HCC neoplastic tissue specimens embedded by paraffin wax which matched with study-required were collected.The modified tissue-cracked assay was used to extract genomic DNA for construction the genomic DNA library of HCC neoplastic tissues.The obtained genomic library quality was evaluated by SmartSpecTM spectrophotometer for nucleic acid and protein detection,TaqMan-MGB-polymerase chain reaction(PCR) and agarose gel electrophoresis assay.Results In this study,2108 HCC neoplastic tissue specimens embedded by paraffin wax were collected according to the selective criterion and the genomic DNA library of HCC neoplasm tissues was successfully constructed.This library was characterized at the concentration of more than(or near) 0.1 μg/μL,the corresponding ratio of A260 to A280 was 1.6～2.0,the real-time quantitative curve of TaqMan-MGB-PCR with ganoid and mono-wave crest,the graph of agarose gel electrophoresis assay presented with unitary,clear,and non-trailing-phenomenon electrophoresis strip.The constructed genomic DNA library presented good quality, and in comparison of the quality to the control DNA group,there was not statistically significant difference(P>0.05).Conclusion The results suggest that the genomic DNA library of HCC neoplasm tissues should be constructed with the paraffin-embedded tissues. |
| Keywords: | paraffin embedding carcinoma,hepatocellular genomic DNA library quality control |
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