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标记RNA与琼脂糖凝胶中的DNA杂交。

Hybridization of labeled RNA to DNA in agarose gels.

作者信息

Shinnick T M, Lund E, Smithies O, Blattner F R

出版信息

Nucleic Acids Res. 1975 Oct;2(10):1911-29. doi: 10.1093/nar/2.10.1911.

DOI:10.1093/nar/2.10.1911
PMID:1103090
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC343556/
Abstract

Specific DNA restriction endonuclease fragments can be identified after electrophoresis in agarose gels by hybridization in the gel (in situ) to radioactive homologous RNA. RNA-DNA hybrids are detected by autoradiography of the gel. Comparison of band patterns of the autoradiogram and the ethidium bromide stained gel allows the identification of the DNA fragment which is complementary to the RNA probe. The technique is rapid, easy and inexpensive. It is sensitive enough to detect individual genes in a mixture of fragments produced by restriction enzyme digestion of complex cellular DNA. We have used this technique to determine which of the Hin III and Eco R1 fragments of phi80d3ilv+su+7 and E. coli DNAs contain the 5S, 16S and 23S ribosomal RNA (rRNA) genes of E. coli.

摘要

特定的DNA限制性内切酶片段在琼脂糖凝胶中电泳后,可通过与放射性同源RNA在凝胶中(原位)杂交来鉴定。通过凝胶放射自显影检测RNA-DNA杂交体。比较放射自显影片和溴化乙锭染色凝胶的条带模式,可鉴定出与RNA探针互补的DNA片段。该技术快速、简便且成本低廉。它灵敏度足够高,能够在复杂细胞DNA经限制性酶消化产生的片段混合物中检测到单个基因。我们已使用该技术来确定phi80d3ilv+su+7和大肠杆菌DNA的Hin III和Eco R1片段中哪些包含大肠杆菌的5S、16S和23S核糖体RNA(rRNA)基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e58/343556/951a13a4acb7/nar00507-0286-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e58/343556/1eb952f18a8d/nar00507-0278-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e58/343556/beddf0223936/nar00507-0280-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e58/343556/b29310487a02/nar00507-0281-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e58/343556/2016a8ddf5f5/nar00507-0284-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e58/343556/951a13a4acb7/nar00507-0286-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e58/343556/1eb952f18a8d/nar00507-0278-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e58/343556/beddf0223936/nar00507-0280-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e58/343556/b29310487a02/nar00507-0281-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e58/343556/2016a8ddf5f5/nar00507-0284-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e58/343556/951a13a4acb7/nar00507-0286-a.jpg

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Plasmids in Yersinia pestis.鼠疫耶尔森氏菌中的质粒

本文引用的文献

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A restriction enzyme from Hemophilus influenzae. II.一种来自流感嗜血杆菌的限制性内切酶。II.
J Mol Biol. 1970 Jul 28;51(2):393-409. doi: 10.1016/0022-2836(70)90150-6.
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Tandem synthesis of the 16S and 23S ribosomal RNA sequences of Escherichia coli.大肠杆菌16S和23S核糖体RNA序列的串联合成
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Deoxyribonucleic acid-ribonucleic acid hybridization. Annealing and quantitative recovery of intact ribosomal ribonucleic acid molecules from hybrids.脱氧核糖核酸-核糖核酸杂交。从杂交体中退火并定量回收完整的核糖体核糖核酸分子。
Infect Immun. 1981 Feb;31(2):839-41. doi: 10.1128/iai.31.2.839-841.1981.
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Polarity of amber mutations in ribosomal protein genes of Escherichia coli.大肠杆菌核糖体蛋白基因中琥珀突变的极性
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Use of randomly cloned DNA fragments for identification of Bacteroides thetaiotaomicron.使用随机克隆的DNA片段鉴定嗜内脏拟杆菌。
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Attenuation regulation in the thr operon of Escherichia coli K-12: molecular cloning and transcription of the controlling region.大肠杆菌K-12苏氨酸操纵子中的衰减调控:控制区域的分子克隆与转录
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Three different mutations in codon 61 of the human N-ras gene detected by synthetic oligonucleotide hybridization.通过合成寡核苷酸杂交检测到人类N-ras基因第61密码子的三种不同突变。
Nucleic Acids Res. 1984 Dec 11;12(23):9155-63. doi: 10.1093/nar/12.23.9155.
8
Detection of sickle cell beta S-globin allele by hybridization with synthetic oligonucleotides.通过与合成寡核苷酸杂交检测镰状细胞βS-珠蛋白等位基因。
Proc Natl Acad Sci U S A. 1983 Jan;80(1):278-82. doi: 10.1073/pnas.80.1.278.
9
Import of proteins into mitochondria: nucleotide sequence of the gene for a 70-kd protein of the yeast mitochondrial outer membrane.蛋白质导入线粒体:酵母线粒体外膜70kd蛋白质基因的核苷酸序列。
EMBO J. 1983;2(12):2169-72. doi: 10.1002/j.1460-2075.1983.tb01718.x.
10
Detection and mapping of homologous, repeated and amplified DNA sequences by DNA renaturation in agarose gels.通过琼脂糖凝胶中的DNA复性检测和定位同源、重复和扩增的DNA序列。
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Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. VIII. The structure of bacteriophage phi 80d-3ilv+su+7, including the mapping of the ribosomal RNA genes.大肠杆菌质粒间序列关系的电子显微镜异源双链研究。VIII. 噬菌体φ80d - 3ilv⁺su⁺7的结构,包括核糖体RNA基因的定位
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6
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J Virol. 1974 Nov;14(5):1235-44. doi: 10.1128/JVI.14.5.1235-1244.1974.
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Proc Natl Acad Sci U S A. 1974 May;71(5):2077-81. doi: 10.1073/pnas.71.5.2077.
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Proc Natl Acad Sci U S A. 1972 Nov;69(11):3448-52. doi: 10.1073/pnas.69.11.3448.