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大鼠内耳组织微量核酸的提取及PCR扩增 被引量:1
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作者 王琼 孔维佳 《上海实验动物科学》 CAS 2002年第2期92-94,共3页
分别用提质粒法 ,SDS-蛋白酶 K法 ,TRIZOL 法提取大鼠内耳组织微量核酸 ,对所提核酸进行紫外吸收光谱分析及 PCR扩增。结果表明 ,提质粒法可制备不含核 DNA的线粒体 DNA;SDS-蛋白酶 K法可制备总 DNA ;TRIZOL法可同时制备总 DNA和总RNA。
关键词 耳聋 动物模型 大鼠 内耳组织 微量核酸 提取 PCR扩增
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新型酸性银染技术在微量核酸检测中的应用 被引量:1
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作者 余虹 石建党 +1 位作者 张维铭 步怀东 《中华病理学杂志》 CAS CSCD 北大核心 1996年第1期48-49,共2页
关键词 微量核酸 检测 银染技术 酸性 基因诊断
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简便有效的微量样品核酸DNA的抽提方法 被引量:1
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作者 张炎 李俊 《生物技术》 CAS CSCD 2000年第5期38-41,共4页
关键词 微量核酸DNA 抽提方法 TWEEN 80 DTT
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微量核酸释放剂提取病毒DNA的使用方法
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《药物生物技术》 CAS 2015年第5期406-406,共1页
一种利用微量核酸释放剂提取生物样本中病毒DNA的方法,属于分子生物学技术领域,特别是涉及利用微量核酸释放剂提取病毒核酸(DNA)的试剂及其使用方法;微量核酸释放剂裂解生物样本(血清或血浆)中病毒DNA同时,佐以促释放剂能更有... 一种利用微量核酸释放剂提取生物样本中病毒DNA的方法,属于分子生物学技术领域,特别是涉及利用微量核酸释放剂提取病毒核酸(DNA)的试剂及其使用方法;微量核酸释放剂裂解生物样本(血清或血浆)中病毒DNA同时,佐以促释放剂能更有效地保证裂解、释放效率;微量核酸释放剂在完全释放核酸的同时,还具有封闭样本中对PCR扩增具有干扰作用的蛋白质、药物及溶血等各种因素物质的功能,避免检测过程的假阴性;样本提取简便、灵敏、快速、准确,避免反复离心、洗脱开盖所引起的污染和核酸丢失,实现PCR提取的“一步法”和“一室化”操作。 展开更多
关键词 病毒DNA 微量核酸 释放剂 提取 分子生物学技术 PCR扩增 生物样本 病毒核酸
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Intrahepatic expression of genes related to metabotropic receptors in chronic hepatitis 被引量:2
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作者 Andrzej Ciesla Maciej Kusmider +6 位作者 Agata Faron-Górecka Marta Dziedzicka-Wasylewska Monika Bocisga-Jasik Danuta Owczarek Irena Cieko-Michalska Dorota Cibor Tomasz Mach 《World Journal of Gastroenterology》 SCIE CAS CSCD 2012年第31期4156-4161,共6页
AIM: To screen for genes related to metabotropic re- ceptors that might be involved in the development of chronic hepatitis. METHODS: Assessment of 20 genes associated with metabotropic receptors was performed in li... AIM: To screen for genes related to metabotropic re- ceptors that might be involved in the development of chronic hepatitis. METHODS: Assessment of 20 genes associated with metabotropic receptors was performed in liver speci- mens obtained by punch biopsy from 12 patients with autoimmune and chronic hepatitis type B and C. For this purpose, a microarray with low integrity grade and with oligonucleotide DNA probes complementary to target transcripts was used. Evaluation of gene expression was performed in relation to transcript level, correlation between samples and grouping of clinical parameters used in chronic hepatitis assessment. Clini- cal markers of chronic hepatitis included alanine and aspartate aminotransferase, ~,-glutamyltranspeptidase, alkaline phosphatase and cholinesterase activity, levels of iron ions, total cholesterol, triglycerides, albumin, glucose, hemoglobin, platelets, histological analysis of inflammatory and necrotic status, fibrosis according to METAVIR score, steatosis, as well as anthropometric body mass index, waist/hip index, percentage of adi- pose tissue and liver size in ultrasound examination. Gender, age, concomitant diseases and drugs were also taken into account. Validation of oligonucleotide microarray gene expression results was done with the use of quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: The highest (0.002 〈 P 〈 0.046) expres- sion among genes encoding main components of metabotropic receptor pathways, such as the a subunit of G-coupled protein, phosphoinositol-dependent pro- tein kinase or arrestin was comparable to that of an- giotensinogen synthesized in the liver. Carcinogenesis suppressor genes, such as chemokine ligand 4, tran- scription factor early growth response protein 1 and lysophosphatidic acid receptor, were characterized by the lowest expression (0.002 〈 P 〈 0.046), while the factor potentially triggering hepatic cancer, transcrip- tion factor JUN-B, had a 20-fold higher expression. The correlation between expression of genes of protein kinases PDPK1, phosphoinositide 3-kinase and protein kinase A (Spearman's coefficient range: 0.762-0.769) confirmed a functional link between these enzymes. Gender (P = 0.0046) and inflammation severity, mea- sured by alanine aminotransferase activity (P = 0.035), were characterized by diverse metabotropic receptor gene expression patterns. The Pearson's coefficient ranging from -0.35 to 0.99 from the results of qRT-PCR and microarray indicated that qRT-PCR had certainlimitations as a validation tool for oligonucleotide mi- croarray studies. CONCLUSION: A microarray-based analysis of hepa- tocyte metabotropic G-protein-related gene expression can reveal the molecular basis of chronic hepatitis. 展开更多
关键词 Metabotropic receptors Gene expression DNA oligonucleotides Quantitative real-time poly-merase chain reaction Chronic hepatitis
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微量COBASAmpliScreen乙肝测试系统筛查供血的乙型肝炎病毒DNA
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作者 杨华松(摘译) 赵国胜(校) 《国际输血及血液学杂志》 CAS 2006年第2期162-162,共1页
关键词 AmpliScreen 乙型肝炎病毒DNA 微量核酸 乙型肝炎病毒(HBV) COBAS 人类免疫缺陷病毒(HIV) 测试系统 丙型肝炎病毒(HCV) 供血 筛查
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