Detection of specific sequences among DNA fragments separated by gel electrophoresis.

  title={Detection of specific sequences among DNA fragments separated by gel electrophoresis.},
  author={Edwin M. Southern},
  journal={Journal of molecular biology},
  volume={98 3},
  • E. Southern
  • Published 1975
  • Biology, Medicine
  • Journal of molecular biology
This paper describes a method of transferring fragments of DNA from agarose gels to cellulose nitrate filters. The fragments can then be hybridized to radioactive RNA and hybrids detected by radioautography or fluorography. The method is illustrated by analyses of restriction fragments complementary to ribosomal RNAs from Escherichia coli and Xenopus laevis , and from several mammals. 
Quantitative electrophoretic transfer of DNA from polyacrylamide or agarose gels to nitrocellulose.
A method for efficient electrophoretic transfer of DNA fragments from polyacrylamide gels to nitrocellulose sheets was developed, allowing this transfer method to be used for DNA ranging from 40 to at least 23,000 bp. Expand
Membrane electrophoresis of DNA
This work has separated double stranded DNA fragments by electrophoresis using a membrane instead of a gel as separating matrix and therefore eliminates the need for a transfer or blotting step. Expand
A rapid two-dimensional fractionation of DNA restriction fragments.
Genomic DNA that has been digested with a restriction endonuclease and fractionated by electrophoresis on an agarose gel can be recovered on glass-fiber filters by a new blotting scheme, useful in the analysis of complex genomes and in the isolation and cloning of particular sequences. Expand
Transfer of small plasmid DNA fragments from polyacrylamide gels onto nitrocellulose paper.
The incorporation into a 7% polyacrylamide gel of a nucleic acid-specific photochemical reagent 4,5',8-trimethylpsoralen at a concentration of 1 mg/dl of acrylamide solution improves both theExpand
Rapid DNA purification for restriction fragment length polymorphism analysis.
This paper describes a method for isolation of DNA from blood samples involving a rapid chemical disintegration of proteins with 8 M urea and with a minimum of exposure to phenol. The DNA is furtherExpand
Direct fractionation of genes by preparative electrophoresis of Bacillus subtilis DNA.
Discontinuous electrophoresis through agarose has been shown to be a satisfactory method for preparation of biologically active restriction fragments from milligram quantities of DNA. The DNA isExpand
Vacuum transfer of DNA to filters for detecting interindividual polymorphism by Southern's blotting hybridization method
The data are provided on the possibility of detecting interindividual polymorphism from the organization of repeated DNA sequences of the alphoid type in human genome on the basis of the Sothern blotting hybridization method. Expand
Spot blot: a hybridization assay for specific DNA sequences in multiple samples.
Small aliquots of DNA-containing samples from gradient or column fractions are spotted onto agarose plates containing ethidium bromide, and the DNA is visualized qualitatively, and transferred to nitrocellulose filters for hybridization. Expand
Southern Blotting and Related DNA Detection Techniques
Southern blotting is a technique for transfer of DNA molecules from an electrophoresis gel to a nitrocellulose or nylon membrane, and is carried out prior to detection of specific molecules byExpand
An easy and efficient procedure for the isolation of pure DNA restriction fragments from agarose gels.
It is shown that the isolated DNA can be used for fragmentation by restriction endonucleases, synthesis of complementary RNA by DNA-dependent RNA polymerase from Escherichia coli and nick translation. Expand


A procedure for the isolation of deoxyribonucleic acid from micro-organisms
A method has been described for the isolation of DNA from micro-organisms which yields stable, biologically active, highly polymerized preparations relatively free from protein and RNA, and Representative samples have been characterized for their thermal stability and sedimentation behaviour. Expand
Hybridization of filters with competitor DNA in the liquid phase in a standard and a micro-assay.
Under appropriate conditions, radioactive RNA (or DNA) can be hybridized with DNA on filters in the presence of denatured DNA in the liquid phase and the method provides means of gaining the same sort of information which is obtained in several steps by differential filter hybridization. Expand
Studies on the cleavage of bacteriophage lambda DNA with EcoRI Restriction endonuclease.
The five EcoRI† restriction sites in bacteriophage lambda DNA have been mapped and the DNA lengths of the EcoRI fragments are in agreement with their electrophoretic mobility on agarose gels but are not in Agreement with their mobilities on polyacrylamide gels. Expand
Molecular hybridization between rat liver deoxyribonucleic acid and complementary ribonucleic acid.
Evidence is presented suggesting that the apparently diminished capacity of the DNA to hybridize is due to loss of hybridized DNA from the membrane filters and it is suggested that the most highly reiterated DNA component is poorly transcribed in vitro. Expand
Amplified ribosomal DNA from Xenopus laevis has heterogeneous spacer lengths.
  • P. Wellauer, R. Reeder, +4 authors I. Dawid
  • Biology, Medicine
  • Proceedings of the National Academy of Sciences of the United States of America
  • 1974
Heterogeneity is present in the DNA amplified from the rDNA of a single nucleolar organizer, and heterogeneity is located in the nontranscribed spacer region which is variable in length, demonstrated by the formation of deletion loops in heteroduplexes made between larger fragments of different lengths. Expand
The determination of the molecular weight of ribonucleic acid by polyacrylamide-gel electrophresis. The effects of changes in conformation.
  • U. Loening
  • Chemistry, Medicine
  • The Biochemical journal
  • 1969
It is concluded that the method provides a satisfactory measurement of molecular weight, which is almost independent of the nucleotide composition of RNA at moderate salt concentrations. Expand
A restriction enzyme from Hemophilus influenzae. I. Purification and general properties.
Abstract Extracts of Hemophilus influenzae strain Rd contain an endonuclease activity which produces a rapid decrease in the specific viscosity of a variety of foreign native DNA's; the specificExpand
A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels.
A simple method for detecting 3H in polyacrylamide gels by scintillation autography (fluorography) using X-ray film, which is ten times more sensitive than conventional autoradiography of isotopes with higher emission energies. Expand
A restriction enzyme from Hemophilus influenzae: II. Base sequence of the recognition site
It is concluded that endonuclease R of H. influenzae recognizes the following specific nucleotide sequence: 5′ … pGpTpPy ¦pPupApCp … 3′ 3′… pCpApPup ¦PypTpGp … 5′ The implications of the twofold rotational symmetry of this sequence are discussed. Expand