Direct analysis of single nucleotide variation in human DNA and RNA using in situ dot hybridization.
Direct analysis of single nucleotide variation in human DNA and RNA using in situ dot hybridization.
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使用原位点杂交直接分析人类 DNA 和 RNA 中的单核苷酸变异。
DOI：
10.1089/dna.1.1989.8.135
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
R. Wallace
中科院分区：
文献类型：
--
作者：
Dan Y. Wu;G. Nozari;M. Schöld;BRENDA J. Conner;R. Wallace
使用寡核苷酸杂交，等位基因之间的单个和多个核苷酸的差异直接在基因组DNA中检测，而无需电泳分离。将DNA固定在琼脂糖凝胶的凹陷处（原位点），并与放射性标记的等位基因特异性寡核苷酸探针杂交。与人类主要组织相容性复合体基因HLA-B27的独特序列区域互补的寡核苷酸仅与来自HLA-B27阳性个体的基因组DNA杂交。合成了与正常人β-珠蛋白基因（β A）和镰状细胞珠蛋白基因（β S）互补的另外两种寡核苷酸。使用竞争性杂交条件，包括存在10倍摩尔过量的与另一个β-珠蛋白等位基因互补的未标记寡核苷酸，来自正常β-珠蛋白基因（β A β A）纯合个体的DNA仅与β A探针杂交，而来自镰状细胞珠蛋白基因（β S β S）纯合个体的DNA仅与镰状细胞基因的探针杂交。正如预期的那样，来自杂合个体的DNA与两种探针结合。用固定在原位点中的总人RNA获得了类似的结果。这种方法可能的应用包括遗传疾病诊断、人群携带者筛查、HLA“DNA”分型以及DNA和RNA序列多态性分析。
Using oligonucleotide hybridization, single and multiple nucleotide differences between alleles were detected directly in genomic DNA without electrophoretic separation. The DNA was immobilized in depressions in an agarose gel (in situ dots) and hybridized with radiolabeled, allele-specific oligonucleotide probes. An oligonucleotide complementary to a unique sequence region of the human major histocompatibility complex gene HLA-B27 only hybridized with genomic DNA from an HLA-B27-positive individual. Two other oligonucleotides complementary to the normal human beta-globin gene (beta A) and to the sickle cell globin gene (beta S) were synthesized. Using competition hybridization conditions which included the presence of a 10-fold molar excess of unlabeled oligonucleotide complementary to the other beta-globin allele, DNA from individuals homozygous for the normal beta-globin gene (beta A beta A) hybridized to the beta A probe exclusively, whereas DNA from individuals homozygous for the sickle cell globin gene (beta S beta S) hybridized only with the probe for the sickle cell gene. As expected, DNA from heterozygous individuals bound to both probes. Similar results were obtained with total human RNA immobilized in in situ dots. Possible applications of this methodology include genetic disease diagnosis, population carrier screening, HLA "DNA" typing, and DNA and RNA sequence polymorphism analysis.
通过 DNA 分析改进镰状突变检测：在产前诊断中的应用。
DOI：
10.1056/nejm198207013070106
发表时间：
1982
期刊：
The New England journal of medicine
影响因子：
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作者：
Orkin,SH;Little,PF;KazazianJr,HH;Boehm,CD
通讯作者：
Boehm,CD
DNA 分析在血红蛋白疾病诊断中的应用。
影响因子：
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作者：
Goossens,M;Kan,YY
通讯作者：
Kan,YY
一种针对镰状细胞性贫血的敏感新产前检测。
DOI：
10.1056/nejm198207013070105
发表时间：
1982
期刊：
The New England journal of medicine
影响因子：
--
作者：
Chang,JC;Kan,YW
通讯作者：
Kan,YW