Nucleic Acids Research, 1985, Vol. 13, No. 20 7207-7221
© 1985
Articles |
Rapid transfer of DNA from agarose gels to nylon membranes
Department of Biochemistry, Faculty of Science, Australian National University Canberra A.C.T. 2601, Australia
Received September 3, 1985. Accepted September 26, 1985.
The unique properties of nylon membranes allow for dramatic improvement in the capillary transfer of DNA restriction fragments from agarose gels (Southern blotting). By using 0.4 M NaOH as the transfer solvent following a short pre-treatment of the gel in acid, DNA is depurinated during transfer. Fragments of all sizes are eluted and retained quantitatively by the membrane; furthermore, the alkaline solvent induces covalent fixation of DNA to the membrane. The saving in time and materials afforded by this simple modification is accompanied by a marked improvement in resolution and a ten-fold increase in sensitivity of subsequent hybridization analyses. In addition, we have found that nylon membrane completely retains native (and denatured) DNA in transfer solvents of low lonic strength (including distilled water), although quantitative elution of DNA from the gel is limited to fragments smaller than 4 Kb. This property can be utilized in the direct electrophoretic transfer of native restriction fragments from polyacrylamide gels. Exposure of DNA to ultraviolet light, either in the gel or following transfer to nylon membrane, reduces its ability to hybridize.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  J. Sambrook and D. W. Russell Southern Blotting: Capillary Transfer of DNA to Membranes CSH Protocols, May 1, 2006; 2006(2): pdb.prot4040 - pdb.prot4040. [Full Text]
|
|
|
|
|
|
J. Sambrook and D. W. Russell Southern Blotting: Simultaneous Transfer of DNA from a Single Agarose Gel to Two Membranes CSH Protocols, May 1, 2006; 2006(2): pdb.prot4043 - pdb.prot4043. [Full Text]
|
|
|
|
|
|
J. Sambrook and D. W. Russell Transfer and Fixation of Denatured RNA to Membranes CSH Protocols, May 1, 2006; 2006(2): pdb.prot4060 - pdb.prot4060. [Full Text]
|
|
|
|
 |
|
 M. Falque, L. Decousset, D. Dervins, A.-M. Jacob, J. Joets, J.-P. Martinant, X. Raffoux, N. Ribiere, C. Ridel, D. Samson, et al. Linkage Mapping of 1454 New Maize Candidate Gene Loci Genetics, August 1, 2005; 170(4): 1957 - 1966. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. M. Li, D. Rotter, S. A. Bonos, W. A. Meyer, and F. C. Belanger Identification of a Gene in the Process of Being Lost from the Genus Agrostis Plant Physiology, August 1, 2005; 138(4): 2386 - 2395. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. G. A. EL-Osta, A. J. Hillier, and M. Dobos Construction of a combined physical and genetic map of the chromosome of Lactobacillus acidophilus ATCC 4356 and characterization of the rRNA operons Microbiology, March 1, 2005; 151(3): 875 - 892. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. G. Kossykh and R. S. Lloyd A DNA Adenine Methyltransferase of Escherichia coli That Is Cell Cycle Regulated and Essential for Viability J. Bacteriol., April 1, 2004; 186(7): 2061 - 2067. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. M. Givens, M.-H. Lin, D. J. Taylor, U. Mechold, J. O. Berry, and V. J. Hernandez Inducible Expression, Enzymatic Activity, and Origin of Higher Plant Homologues of Bacterial RelA/SpoT Stress Proteins in Nicotiana tabacum J. Biol. Chem., February 27, 2004; 279(9): 7495 - 7504. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Venkatesh, P. Kumar, P. S. M. Krishna, R. Manjunath, and U. Varshney Importance of Uracil DNA Glycosylase in Pseudomonas aeruginosa and Mycobacterium smegmatis, G+C-rich Bacteria, in Mutation Prevention, Tolerance to Acidified Nitrite, and Endurance in Mouse Macrophages J. Biol. Chem., June 27, 2003; 278(27): 24350 - 24358. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. K. Dineshkumar, S. Thanedar, C. Subbulakshmi, and U. Varshney An unexpected absence of queuosine modification in the tRNAs of an Escherichia coli B strain Microbiology, December 1, 2002; 148(12): 3779 - 3787. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Moy, H. M. Li, R. Sullivan, J. F. White Jr., and F. C. Belanger Endophytic Fungal beta -1,6-Glucanase Expression in the Infected Host Grass Plant Physiology, November 1, 2002; 130(3): 1298 - 1308. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. T. O'Connell, J. S. Tomlinson, A. A. Roberts, K. F. McGonigle, and S. H. Barsky Pseudomyxoma Peritonei Is a Disease of MUC2-Expressing Goblet Cells Am. J. Pathol., August 1, 2002; 161(2): 551 - 564. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Chong, J. Kontaraki, C. Bonifer, and A. D. Riggs A Functional Chromatin Domain Does Not Resist X Chromosome Inactivation: Silencing of cLys Correlates with Methylation of a Dual Promoter-Replication Origin Mol. Cell. Biol., July 1, 2002; 22(13): 4667 - 4676. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Plohl, E. Prats, A. Martinez-Lage, A. Gonzalez-Tizon, J. Mendez, and L. Cornudella Telomeric Localization of the Vertebrate-type Hexamer Repeat, (TTAGGG)n, in the Wedgeshell Clam Donax trunculus and Other Marine Invertebrate Genomes J. Biol. Chem., May 24, 2002; 277(22): 19839 - 19846. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. A. Lockman, R. A. Gillespie, B. D. Baker, and E. Shakhnovich Yersinia pseudotuberculosis Produces a Cytotoxic Necrotizing Factor Infect. Immun., May 1, 2002; 70(5): 2708 - 2714. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. B. Tennyson, N. Ebran, A. E. Herrera, and J. E. Lindsley A Novel Selection System for Chromosome Translocations in Saccharomyces cerevisiae Genetics, April 1, 2002; 160(4): 1363 - 1373. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. M. Karrer and T. A. VanNuland Methylation of adenine in the nuclear DNA of Tetrahymena is internucleosomal and independent of histone H1 Nucleic Acids Res., March 15, 2002; 30(6): 1364 - 1370. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Watanabe, R. Zeng, Y. Aoki, T. Itoh, and K.-i. Arai Initiation of polyoma virus origin-dependent DNA replication through STAT5 activation by human granulocyte-macrophage colony-stimulating factor Blood, March 1, 2001; 97(5): 1266 - 1273. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. T. Villar, R. L. Hirschberg, and M. R. Schaefer Role of the Eikenella corrodens pilA Locus in Pilus Function and Phase Variation J. Bacteriol., January 1, 2001; 183(1): 55 - 62. [Abstract] [Full Text]
|
|
|
|
|
|
D.-S. Sheu, Y.-T. Wang, and C.-Y. Lee Rapid detection of polyhydroxyalkanoate-accumulating bacteria isolated from the environment by colony PCR Microbiology, August 1, 2000; 146(8): 2019 - 2025. [Abstract] [Full Text]
|
|
|
|
|
|
W. R. Zuckert and A. G. Barbour Stability of Borrelia burgdorferi bdr Loci In Vitro and In Vivo Infect. Immun., March 1, 2000; 68(3): 1727 - 1730. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. M. Karrer and T. A. VanNuland Nucleosome Positioning Is Independent of Histone H1 in Vivo J. Biol. Chem., November 12, 1999; 274(46): 33020 - 33024. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. J. Morrow, J. E. Graham, and R. Curtiss III Genomic Subtractive Hybridization and Selective Capture of Transcribed Sequences Identify a Novel Salmonella typhimurium Fimbrial Operon and Putative Transcriptional Regulator That Are Absent from the Salmonella typhi Genome Infect. Immun., October 1, 1999; 67(10): 5106 - 5116. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Y. Yount, J. Yuan, A. Tarver, T. Castro, G. Diamond, P. A. Tran, J. N. Levy, C. McCullough, J. S. Cullor, C. L. Bevins, et al. Cloning and Expression of Bovine Neutrophil beta -Defensins. BIOSYNTHETIC PROFILE DURING NEUTROPHILIC MATURATION AND LOCALIZATION OF MATURE PEPTIDE TO NOVEL CYTOPLASMIC DENSE GRANULES J. Biol. Chem., September 10, 1999; 274(37): 26249 - 26258. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. G. Stathakis, D. Y. Burton, W. E. McIvor, S. Krishnakumar, T. R. F. Wright, and J. M. O'Donnell The Catecholamines up (Catsup) Protein of Drosophila melanogaster Functions as a Negative Regulator of Tyrosine Hydroxylase Activity Genetics, September 1, 1999; 153(1): 361 - 382. [Abstract] [Full Text]
|
|
|
|
 |
|
 M. Fuchs, P. Hübert, J. Detterer, and H.-J. Rziha Detection of Bovine Herpesvirus Type 1 in Blood from Naturally Infected Cattle by Using a Sensitive PCR That Discriminates between Wild-Type Virus and Virus Lacking Glycoprotein E J. Clin. Microbiol., August 1, 1999; 37(8): 2498 - 2507. [Abstract] [Full Text]
|
|
|
|
|
|
M. T. Villar, J. T. Helber, B. Hood, M. R. Schaefer, and R. L. Hirschberg Eikenella corrodens Phase Variation Involves a Posttranslational Event in Pilus Formation J. Bacteriol., July 15, 1999; 181(14): 4154 - 4160. [Abstract] [Full Text]
|
|
|
|
 |
|
 K. E. Wommack, J. Ravel, R. T. Hill, and R. R. Colwell Hybridization Analysis of Chesapeake Bay Virioplankton Appl. Envir. Microbiol., January 1, 1999; 65(1): 241 - 250. [Abstract] [Full Text]
|
|
|
|
 |
|
 B. Essigmann, S. Guler, R. A. Narang, D. Linke, and C. Benning Phosphate availability affects the thylakoid lipid composition and the expression of SQD1, a gene required for sulfolipid biosynthesis in Arabidopsis thaliana PNAS, February 17, 1998; 95(4): 1950 - 1955. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Faustino, L. Osorio-Almeida, L. Romao, J. Barbot, B. Fernandes, B. Justica, and J. Lavinha Dominantly Transmitted beta -Thalassemia Arising From the Production of Several Aberrant mRNA Species and One Abnormal Peptide Blood, January 15, 1998; 91(2): 685 - 690. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Castle, D. Speranzini, N. Rghei, G. Alm, D. Rinker, and J. Bissett Morphological and Molecular Identification of Trichoderma Isolates on North American Mushroom Farms Appl. Envir. Microbiol., January 1, 1998; 64(1): 133 - 137. [Abstract] [Full Text]
|
|
|
|
|
|
A. Bito, M. Haider, I. Hadler, and M. Breitenbach Identification and Phenotypic Analysis of Two Glyoxalase II Encoding Genes from Saccharomyces cerevisiae, GLO2 and GLO4, and Intracellular Localization of the Corresponding Proteins J. Biol. Chem., August 22, 1997; 272(34): 21509 - 21519. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I Guenal, Y Risler, and B Mignotte Down-regulation of actin genes precedes microfilament network disruption and actin cleavage during p53-mediated apoptosis J. Cell Sci., January 2, 1997; 110(4): 489 - 495. [Abstract] [PDF]
|
|
|
|
|
|
S. Güler, A. Seeliger, H. Härtel, G. Renger, and C. Benning A Null Mutant of Synechococcus sp. PCC7942 Deficient in the Sulfolipid Sulfoquinovosyl Diacylglycerol J. Biol. Chem., March 29, 1996; 271(13): 7501 - 7507. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. M. Owczarek, M. J. Layton, L. G. Robb, N. A. Nicola, and C. G. Begley Molecular Basis of the Soluble and Membrane-bound Forms of the Murine Leukemia Inhibitory Factor Receptor alpha-Chain J. Biol. Chem., March 8, 1996; 271(10): 5495 - 5504. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. A. Wetsel, J. Kulics, M.-L. Lokki, P. Kiepiela, H. Akama, C. A. C. Johnson, P. Densen, and H. R. Colten Type II Human Complement C2 Deficiency J. Biol. Chem., March 8, 1996; 271(10): 5824 - 5831. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Cui and D. E. Vance Expression of Phosphatidylethanolamine N-Methyltransferase-2 Is Markedly Enhanced in Long Term Choline-deficient Rats J. Biol. Chem., February 2, 1996; 271(5): 2839 - 2843. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Tominaga, T. Tominaga, A. Miwa, and Y. Okada Volume-sensitive Chloride Channel Activity Does Not Depend on Endogenous P-glycoprotein J. Biol. Chem., November 17, 1995; 270(46): 27887 - 27893. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Watanabe, Y. Ito, A. Miyajima, and K.-i. Arai Granulocyte Macrophage-Colony Stimulating Factor-dependent Replication of Polyoma Virus Replicon in Hematopoietic Cells J. Biol. Chem., April 21, 1995; 270(16): 9615 - 9621. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M Murphy, K Reid, M. Brown, and P. Bartlett Involvement of leukemia inhibitory factor and nerve growth factor in the development of dorsal root ganglion neurons Development, January 3, 1993; 117(3): 1173 - 1182. [Abstract] [PDF]
|
|
|
|
 |
|
 J Yourno A method for nested PCR with single closed reaction tubes. Genome Res., August 1, 1992; 2(1): 60 - 65. [Abstract] [PDF]
|
|
|
|
 |
|
 D Sidransky, T Tokino, Hamilton SR, K. Kinzler, B Levin, P Frost, and B Vogelstein Identification of ras oncogene mutations in the stool of patients with curable colorectal tumors Science, April 3, 1992; 256(5053): 102 - 105. [Abstract] [PDF]
|
|
|
|
 |
|
 W J Buikema and R Haselkorn Characterization of a gene controlling heterocyst differentiation in the cyanobacterium Anabaena 7120. Genes & Dev., February 1, 1991; 5(2): 321 - 330. [Abstract] [PDF]
|
|
|
|
|
|
C A Brenner, R R Adler, D A Rappolee, R A Pedersen, and Z Werb Genes for extracellular-matrix-degrading metalloproteinases and their inhibitor, TIMP, are expressed during early mammalian development. Genes & Dev., June 1, 1989; 3(6): 848 - 859. [Abstract] [PDF]
|
|
|
|
|
|
J A Banks, P Masson, and N Fedoroff Molecular mechanisms in the developmental regulation of the maize Suppressor-mutator transposable element. Genes & Dev., November 1, 1988; 2(11): 1364 - 1380. [Abstract] [PDF]
|
|
|
|
|
|
G. Schellenberg, T. Bird, E. Wijsman, D. Moore, M Boehnke, E. Bryant, T. Lampe, D Nochlin, S. Sumi, S. Deeb, et al. Absence of linkage of chromosome 21q21 markers to familial Alzheimer's disease Science, September 16, 1988; 241(4872): 1507 - 1510. [Abstract] [PDF]
|
|
|
|
|
|
B I Osborne and L Guarente Transcription by RNA polymerase II induces changes of DNA topology in yeast. Genes & Dev., June 1, 1988; 2(6): 766 - 772. [Abstract] [PDF]
|
|
|
|
|
|
R. Saiki, D. Gelfand, S Stoffel, S. Scharf, R Higuchi, G. Horn, K. Mullis, and H. Erlich Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase Science, January 29, 1988; 239(4839): 487 - 491. [Abstract] [PDF]
|
|
|
|
|
|
M Leppert, M Dobbs, P Scambler, P O'Connell, Y Nakamura, D Stauffer, S Woodward, R Burt, J Hughes, E Gardner, et al. The gene for familial polyposis coli maps to the long arm of chromosome 5 Science, December 4, 1987; 238(4832): 1411 - 1413. [Abstract] [PDF]
|
|
|
|
|
|
D Goldgaber, M. Lerman, O. McBride, U Saffiotti, and D. Gajdusek Characterization and chromosomal localization of a cDNA encoding brain amyloid of Alzheimer's disease Science, February 20, 1987; 235(4791): 877 - 880. [Abstract] [PDF]
|
|
|
|
|
|
A. Mason, S. Pitts, K Nikolics, E Szonyi, J. Wilcox, P. Seeburg, and T. Stewart The hypogonadal mouse: reproductive functions restored by gene therapy Science, December 12, 1986; 234(4782): 1372 - 1378. [Abstract] [PDF]
|
|
|
|
|
|
C Moritz and W. Brown Tandem duplication of D-loop and ribosomal RNA sequences in lizard mitochondrial DNA Science, September 26, 1986; 233(4771): 1425 - 1427. [Abstract] [PDF]
|
|
|
|
|
|
T. J. Kotti, K. Savolainen, H. M. Helander, A. Yagi, D. K. Novikov, N. Kalkkinen, E. Conzelmann, J. K. Hiltunen, and W. Schmitz In Mouse alpha -Methylacyl-CoA Racemase, the Same Gene Product Is Simultaneously Located in Mitochondria and Peroxisomes J. Biol. Chem., June 30, 2000; 275(27): 20887 - 20895. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.-P. Sim and L. F. Liu Nucleolytic Cleavage of the Mixed Lineage Leukemia Breakpoint Cluster Region during Apoptosis J. Biol. Chem., August 17, 2001; 276(34): 31590 - 31595. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. M. Klug and C. Benning Two enzymes of diacylglyceryl-O-4'-(N,N,N,-trimethyl)homoserine biosynthesis are encoded by btaA and btaB in the purple bacterium Rhodobacter sphaeroides PNAS, May 8, 2001; 98(10): 5910 - 5915. [Abstract] [Full Text] [PDF]
|
|