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DIG-System - Literature
Filter Hybridization
Molecular Biology
Page
I.
General papers
II.
Methods
4
1. Labeling techniques
A. DNA probes
a. Random primed labeling
b. Nick translation
c. cDNA synthesis
d. PCR
B. RNA probes
5
6
6
9
C. Oligonucleotide probes
a. 3'Endlabeling
b. 3'Tailing
c. Chemical synthesis (DIG-X-NHS ester)
10
10
10
2. Detection techniques
A. BCIP/NBT detection
10
B. Chemiluminescent detection
11
C. Multicolor detection
11
-2Page
III.
Applications
1. Blots
1.1 Dot blot/slot blot
11
1.1.2 HLA Typing
20
1.2 Southern transfer
A. DNA probes
a. Random primed labeling
b. Nick translation
c. cDNA synthesis
d. PCR
B. RNA probes
21
27
28
28
31
C. Oligonucleotide probes
a. 3'Endlabeling
b. 3'Tailing
c. Chemical synthesis
31
31
33
1.2.1 Special applications of Southern blotting
A. Fingerprinting
33
B. Genome mapping
34
C. RFLP / AFLP analysis
34
1.3 Northern analysis
34
1.4 Colony hybridization
39
1.5 Plaque hybridization
42
-3Page
2. Hybridization in the gel or liquid
42
3. Recombination in vitro
43
4. Assays in Microtitration Wells
43
5. Non-radioactive sequencing
46
6. Non-radioactive RNase Protection Assay
46
7. Non-radioactive Gel Shift Assay
47
8. Protein-RNA Binding
48
9. Nuclear Runoff Transcription Analysis
48
10. Differential Display
48
11. Miscellaneous
48
-4-
I. General papers
Diamandis, E.P.; Hassapoglidou, S.; Bean, C.C. (1993) Evaluation of nonisotopic labeling and
detection techniques for nucleic acid hybridization. J. Clin. Lab. Anal. 7, 174-179.
Düring, K. (1993) Non-radioactive detection methods for nucleic acids separated by
electrophoresis. J. Chromat. 618, 105-131.
Düring, K. (1991) Ultrasensitive chemiluminescent and colorigenic detection of DNA, RNA and
protein in plant molecular biology. Anal. Biochem. 196, 433-438.
Ferguson-Smith, M.A. (1991) Invited Editorial: Putting the genetics back into cytogenetics. Am.
J. Hum. Genet. 48, 179-182.
Guesdon, J.-L. (1992) Immunoenzymatic techniques applied to the specific detection of nucleic
acids. J. Immunol. Meth. 150, 33-49.
Höltke, H.-J. (1992) Nonradioactive labeling of nucleic acids with digoxigenin and detection by
colour, fluorescence and luminescence. ABCLAI 50 (6-7), 496.
Höltke, H.-J.; Ankenbauer, W.; Mühlegger, K.; Rein, R.; Sagner, G.; Seibl, R.; Walter, T. (1995)
The Digoxigenin (DIG) system for non-radioactive labelling and detection of nucleic acids - an
overview. Cell. Mol. Biol. 41 (7), 883-905.
Höltke, H.-J.; Sagner, G.; Kessler, C.; Schmitz, G. (1992) Chemiluminescent detection of
digoxigenin-labeled nucleic acids: Optimization of the detection protocol. ABCLAI 50 (6-7), 496.
Höltke, H.-J.; Seibl, R.; Burg, J.; Mühlegger, K.; Kessler, C. (1990) Non-radioactive labeling and
detection of nucleic acids: II. Optimization of the digoxigenin system. Biol. Chem. Hoppe-Seyler
371, 929-938.
Höltke, H.-J.; Seibl, R.; Burg, J.; Mühlegger, K.; Mattes, R.; Kessler, C. (1988) Non-radioactive
HighSens DNA labeling and detection system (Digoxigenin: anti-digoxigenin based ELISA
principle). Fresenius Z. Anal. Chem. 330, 377-378.
Hughes, J.R., Evans, M.F., Levy, E.R. (1995) Non-isotopic methods in molecular biology.
Oxford: IRL Pr. at Oxford Univ. Pr., 145-182.
Kessler, C. (1990) The digoxigenin system: principle and applications of the novel non-radioactive
DNA labeling and detection system. BioTechnology Intern. 183-194.
Kessler, C. (1990) The digoxigenin: anti-digoxigenin (DIG) technology - a survey on the concept
and realization of a novel bioanalytic indicator system. Mol. Cell. Probes 5, 161-205.
Kessler, C.; Höltke, H.-J.; Seibl, R.; Burg, J.; Mühlegger, K. (1990) Non-radioactive labeling and
detection of nucleic acids: I. A novel DNA labeling and detection system based on digoxigenin:
anti-digoxigenin ELISA principle. Biol. Chem. Hoppe-Seyler 371, 917-927.
-5Kühtz, I.; Gayer-Herkert, G.; Hollstein, I.; Kessler, C. (1991) Digoxigenin-Prinzip und Anwendungen der nichtradioaktiven Markierung und Nachweis von Biomolekülen. BIOforum 14, 100-104.
Martin, R.; Hoover, C.; Grimme, S.; Grogan, C.; Höltke, J.; Kessler, C. (1990) A highly sensitive,
non-radioactive DNA labeling and detection system. BioTechniques 9 (6), 762-768.
McCreery, T. (1997) Digoxigenin Labeling. Mol. Biotechnol. Vol. 7, 121-124
Mitchell, B.S.; Dhami, D.; Schumacher, U. (1992) In situ hybridisation: a review of
methodologies and applications in the biomedical sciences. Med. Lab. Sci. 49, 107-118.
Mühlegger, K.; Huber, E.; v. d. Eltz, H.; Rüger, R.; Kessler, C. (1990) Non-radioactive labeling and
detection of nucleic acids: IV. Synthesis and properties of digoxigenin-modified 2'-deoxyuridine-5'-triphosphates and a photoactivatable analog of digoxigenin (photodigoxigenin). Biol. Chem.
Hoppe-Seyler 371, 953-965.
Mühlegger, K.; Batz, H.-G.; Böhm, S.; v.d. Eltz, H.; Höltke, H.-J.; Kessler, C. (1989) Synthesis and
use of new digoxigenin-labeled nucleotides in non-radioactive labeling and detection of nucleic
acids. Nucleosides & Nucleotides 8 (586), 1161-1163.
Reischl, U.; Mayer, J. (1993) Moderne Methoden der Nukleinsäure-Diagnostik. Lab. med. 17,
456-464.
Sand, T. (1990) New labels; their importance in current and future immunoassays. 22nd Nordic
Congress in Clinical Chemistry, Trondheim, Norway, June 25-28 1990, Scandin. J. Clin. Lab.
Invest. 50, supplement 202.
Seibl, R.; Höltke, H.-J.; Rüger, R.; Meindl, A.; Zachau, H.G.; Raßhofer, R.; Roggendorf, M.;
Wolf, H.; Arnold, N.; Wienberg, J.; Kessler, C. (1990) Non-radioactive labeling and detection of
nucleic acids: III. Applications of the digoxigenin system. Biol. Chem. Hoppe-Seyler 371, 939-951.
Seibl, R.; Höltke, J.; Burg, J.; Mühlegger, K.; Mattes, R.; Kessler, C. (1988) Non-radioactive
HighSens detection of nucleic acids. A novel highly sensitive DNA labeling and detection system
based on digoxigenin: anti-digoxigenin enzyme-linked immuno-sorbent assay (ELISA). Fresenius
Z. Anal. Chem. 330, 305-306.
Thomas, A. (1994) Nonradioactive digoxigenin-labeled DNA probes. Meth. Pract. Labort.
Bacteriol., 107-121.
II. Methods
1. Labeling techniques
A. DNA probes
a. Random primed labeling
Corti, A.; Tripputi, P.; Cassani, G. (1991) A rapid method for monitoring DNA labeling reactions
with haptens. J. Immunol. Method 134, 81-86.
McCreery, T.; Helentjaris, T. (1994) Production of DNA Hybridization Probes with DigoxigeninModified Nucleotides by Random Hexanucleotide Priming. Methods in Mol. Biol., Vol. 28, 73-76.
Rost, A.-K.; Köhler, T.; Heilmann, S.; Lehmann, J.; Remke, H.; Rotzsch, W. (1995) A rapid and
simple method to prepare digoxigenin-labeled DNA-probes by using PCR-generated DNAfragments. Eur. J. Clin. Chem. Clin. Biochem. 33 (4), A59.
Zhan, J., Fahimi, H.D., Vöelkl, A. (1997) Sensitive nonradioactive Dot Blot/Ribonuclease
protection assay for quantitative determination of mRNA. BioTechniques 22, 500-505.
Zhang, Z.S.; Chen, S.Z.; Lin, Z.; Wang, R.Z. (1991) Detection of plasmodium falciparum
infection in humans by (DIG)-DNA probe. Chin. J. Parasitol. Parasit. 9 (4), 287-289.
-6c. cDNA synthesis
Soldo, L.; Mangano, G.; Milanese, C. (1992) Synthesis of digoxigenin-labeled cDNA as a test for
mRNA integrity. BioTechniques 13 (6), 876-879.
d. PCR
(1991) Producing single-stranded DNA probes with the Taq DNA Polymerase: A high yield
protocol. BioTechniques 10 (1), 35-38.
Akin, A.; Wu, C.C.; Lin, T.L.; Keirs, R.W. (1993) Chemiluminescent detection of infectious bursal
disease virus with a PCR-generated nonradiolabeled probe. J. Vet. Diagn. Invest. 5, 166-173.
Alsaadi, S.A. (1993) Preferential hybridization: amplification hybridization signals with nested
probes. BioTechniques 15 (1), 96-99.
An, S.F.; Franklin, D.; Fleming, K.A. (1992) Generation of digoxigenin-labelled double-stranded
and single-stranded probes using the polymerase chain reaction. Mol. Cell. Probes 6, 193-200.
Aschoff, J.M.; Lazarus, D.; Fanburg, B.L.; Lanzillo, J.J. (1994) Relative quantification of
angiotensin-converting enzyme mRNA in human smooth muscle cells, monocytes, and
lymphocytes by the polymerase chain reaction. Anal. Biochem. 219, 218-223.
Batt, C.A.; Wagner, P.; Wiedmann, M.; Luo, J.; Gilbert, R. (1994) Detection of bovine leukocyte
adhesion deficiency by nonisotopic ligase chain reaction. Animal Gen. 25, 95-98.
Besnard, N.C.; Andre, P.M. (1994) Automated quantitative determination of hepatitis C virus
viremia by reverse transcription-PCR. J. Clin. Microbiol. 32 (8), 1887-1893.
Chenal, V.; Griffais, R. (1994) Chemiluminescent and colorimetric detection of a fluoresceinlabelled probe and digoxigenin-labeled probe after a single hybridization step. Mol. Cell. Probes.
8, 401-407.
Durigon, E.L.; Erdmann, D.D.; Anderson, B.C.; Holloway, B.P.; Anderson, L.J. (1994)
Immunochemiluminescent southern blot assay for polymerase chain reaction detection of human
parvovirus. Mol. Cell. Probes 8, 199-204.
Emilsson, V.; Liu, Y.-L.; Cawthorne, M.A.; Morton, N.M.; Davenport, M. (1997) Expression of the
functional leptin receptor mRNA in pancreatic islets and direct inhibitory action of leptin on
insulin secretion. Diabetis, Vol. 46, February 1997 No.2, 313-316.
Erb, R.W.; Wagner-Döbler, I. (1993) Detection of polychlorinated biphenyl degradation genes in
polluted sediments by direct DNA extraction and polymerase chain reaction. Appl. Environ.
Microbiol. 59 (12), 4065-4073.
Figueroa, J.V.; Alvarez, J.A.; Ramos, J.A.; Vega, C.A.; Buening, G.M. (1993) Use of multiplex
polymerase chain reaction-based assay to conduct epidemiological studies on bovine
hemoparasites in Mexico. Revue Elev. Med. vet. Pays trop. 46 (1-2), 71-75.
Finckh, U.; Lingenfelter, P.A.; Myerson, D. (1991) Producing single-stranded DNA probes with the
Taq DNA polymerase: a high yield protocol. BioTechniques 10 (1), 35-39.
Gabellec, M.; Griffais, R.; Fillion, G. Haour, F. (1997) Analysis of brain mRNA by reverse
trancription-polymerase chain reaction and hybridization with digoxigenin-labeled DNA probe.
Brain Research Protocols 1, 145-151.
Gissing, G.J.; Crease, T.J.; Edie, J. (1997) Optimized PCR-labeling and hybridization conditions
result in extremely fast, high quality, chemiluminescent DNA fingerprints. Biochemica No.1, 2324.
-7Gonzalez, N.; Galindo, I.; Guevara, P.; Novak, E.; Scorza, J.V.; Anez, N.; DaSilveira, J.F.;
Ramirez, J.L. (1994) Identification and detection of Trypanosoma cruzi by using a DNA
amplification fingerprint obtained from the ribosomal intergenic spacer. J. Clin. Microbiol. 32 (1),
153-158.
Griffais, R.; Andre, P.M.; Thibon, M. (1990) Synthesis of digoxigenin-labeled DNA probe by
Polymerase Chain Reaction: Application to Epstein-barr virus and chlamydia trachomatis. Res.
Virol. 141, 331-335.
Holmstrom, K.; Rossen, L.; Rasmussen, O.F. (1993) A highly sensitive and fast nonradioactive
method for detection of Polymerase Chain Reaction products. Anal. Biochem. 209, 278-283.
Ibrahim, A.; Liesack, W.; Stackebrandt, E. (1992) Differentiation between pathogenic and nonpathogenic Yersinia enterocolitica strains by colony hybridization with a PCR-mediated
digoxigenin-dUTP-labelled probe. Mol. Cell. Probes 6, 163-171.
Jackson, M.P. (1991) Detection of shiga toxin-producing shigella dysenteriae type 1 and
escherichia coli by using Polymerase Chain Reaction with incorporation of digoxigenin-11-dUTP.
J. Clin. Microbiol. 29 (9), 1910-1914.
Kell, W.M.; LeClaire, R.D.; Schmidt, R.; Langford, M. (1995) A rapid immunochemiluminescence
RT-PCR method for temporal measurement of induced nitric oxide synthase mRNA in
macrophage and endothelial cells. Meth. Mol. Cell Biol. 5, 105-111.
Kessler, H.H.; Pierer, K.; Stünzner, D.; Kleinhappl, B.; Pschaid, A.; Marth, E. (1993) Evaluation
of the Polymerase Chain Reaction for detection of herpesviridae in a transplantation program.
Alpe Adria J. Medicine XLVIII, 253-264.
Kessler, H.H.; Pierer, K.; Weber, B.; Sakrauski, A.; Santner, B.; Stuenzner, D.; Gergely, E.;
Marth, E. (1994) Detection of herpes simplex virus DNA from cerebrospinal fluid by PCR and a
rapid, nonradioactive hybridization technique. J. Clin. Microbiol. 32 (8), 1881-1886
Konat, G.; Laszkiewicz, I.; Bednarczuk, T.; Kanoh, M.; Wiggins, R.C. (1991) Generation of
radioactive and nonradioactive ssDNA hybridization probes by polymerase chain reaction.
Technique 3 (2), 64-68.
Lanzillo, J.L. (1991) Chemiluminescent nucleic acid detection with Digoxigenin-labeled probes: a
model system with probes for angiotensin converting enzyme which detect less than one attomole
of target DNA. Anal. Biochem. 194, 45-53.
Lanzillo, J.L. (1990) Preparation of Digoxigenin-labeled probes by the Polymerase Chain Reaction.
BioTechniques 8 (6), 621-622.
Li, S.-R.; Forster, L.; Änggard, E.; Ferns, G. (1994) The effects of LPS and probucol on
interleukin 1 (IL-1) and platelet-derived growth factor (PDGF) gene expression in the human
monocytic cell line U-937. Bioch. Bioph. Acta 1225, 271-274.
Liesack, W.; Menke, M.A.O.H.; Stackebrandt, E. (1990) Rapid generation of vector-free
digoxygenin-dUTP labeled probes for nonradioactive hybridization using the Polymerase Chain
Reaction (PCR) method. System. Appl. Microbiol. 13, 255 -256.
Lion, T.; Haas, O.A. (1990) Nonradioactive labeling of probe with Digoxigenin by Polymerase
Chain Reaction. Anal. Biochem. 188, 335-337.
Lion, T.; Prischl, F.; Haas, O.A.; Pont, J.; Schwarzmeier, J. (1991) Non-radioactive detection of
the rearranged BCR/ABL sequences amplified by Polymerase Chain Reaction. Leukemia 5 (2),
156-159.
Ludwig, W.; Dorn, S.; Springer, N.; Kirchhof, G.; Schleifer, K.-H. (1994) PCR-based preparation of
23S rRNA-targeted group-specific polynucleotide probes. Appl. Environm. Microbiol. 60 (9), 32363244.
Martin, R.K.; Archer, K.T.; Tuttle, R.M. (1994) Detection of ptc in archival formalin-fixed, paraffinembedded tissues. Diagn. Mol. Pathol. 3 (4), 233-239.
-8Matthes, T.; Kindler, V.; Zubler, R.H. (1994) Semiquantitative, nonradioactive RT-PCR detection
of immunoglobulin mRNA in human B cells and plasma cells. DNA Cell Biol. 13 (4), 429-436.
Miyamoto, K.; Tomita, N.; Ohtsuki, Y; Kitajima, Ko-ichi. (1990) Detection of provirus in an HTLV-II
producer CD8 + T cell line by Polymerase Chain Reaction combined with Digoxigenin - ELISA
method. Jpn. J. Cancer Res. 81, 313- 316.
Moulay, S.; Zientara, S.; SailleauC.; Cruciere, C. (1995) Detection of african horse sickness
viruses by dot-blot hybridization using a digoxigenin-labeled probe. Mol. Cell. Probes 9.
Murthy, V.K.; Dibbern, K.M.; Campbell, D.A. (1992) PCR amplification of mini-exon genes
differentiates Trypanosoma cruzi from Trypanosoma rangeli. Mol. Cel. Probes 6, 237-243.
Nagai, M.; Nemoto, T.; Ota, M. (1994) Mobility of digoxigenin-dUTP polymerase chain reaction
products and detection of single-stranded products in agarose electrophoresis. Anal. Biochem.
221, 208-210.
Nevinny-Stickel, C.; Bettinotti, M. d. l. P.; Andreas, A.; Hinzpeter, M.; Mühlegger, K.; Schmitz, G.;
Albert, E.D. (1990) Non-radioactive HLA class II typing using Polymerase Chain Reaction and
digoxigenin labeled oligonucleotide probes. Human Immunol. 31, 7 - 13.
Nevinny-Stickel, C.; Hinzpeter, M.; Andreas, A.; Albert, E.D. (1991) Non-Radioactive oligotyping
for HLA-DR1-DRw10 using polymerase chain reaction, digoxigenin-labeled oligonucleotides and
chemiluminescence detection. Europ. J. Immunogen. 18, 323-332.
Powell, E.E.; Kroon, P.A. (1992) Measurement of mRNA by quantitative PCR with a nonradioactive label. J. Lipid Res. 33, 609-614.
Preus, H.R.; Russell, D.T. (1994) Use of nonradioactive genetic probe identified, synthesized,
and labeled in the polymerase chain reaction. Scand. J. Dent. Res. 102, 161-167.
Rademaker, C.M.A.; Wolfhagen, M.J.H.M.; Jansze, M.; Oteman, M.; Fluit, A.C.; Glerum, J.H.;
Verhoef, J. (1992) Digoxigenin labelled DNA probes for rapid detection of enteroxigenic,
enteropathogenic and Vero cytotoxin producing Escherichia coli in faecal samples. J. Microbiol.
Meth. 15, 121-127.
Reischl, U.; Rüger, R.; Kessler, C. (1993) Nonradioactive labeling of polymerase chain reaction
products. Meth. Mol. Biol. 15, 51-62.
Reischl, U.; Rüger, R.; Kessler, C. (1994) Nonradioactive labeling and high-sensitive detection of
PCR products. Mol. Biotechn. 1, 229-240.
Reischl, U.; Kochanowski, B. (1995) Quantitative PCR. Mol. Biotechnol. 3, 55-71.
Rettig-Emanual, J. (1991) Simple and efficient system for synthesis of non-radioactive nucleic and
hybridization probes using PCR. Nucleic Acids Res. 19 (10), 2790.
Rodriguez-Frias, F.; Arranz, J.A.; Buti, M.; Esteban, R.; Jardi, R. (1994) Synthesis of a nonradioactive hepatitis B virus DNA probe from human serum by polymerase chain reaction. Eur.
J. Clin. Chem. Clin. Biochem. 32, 355-359.
Rüger, R.; Höltke, H.-J.; Reischl, U.; Sagner, G.; Kessler, C. (1991) Use of Polymerase Chain
Reaction for non-radioactive labelling of specific DNA sequences with digoxigenin. BTF 5,
Advances Mol. Gen. 3, 277-280.
Rüger, R.; Höltke, H.-J.; Reischl, U.; Sagner, G.; Kessler, C. (1990) Use of the Polymerase Chain
Reaction for labeling specific DNA sequences with Digoxigenin. J. Clin. Chem. Clin. Biochem. 28,
566.
Schraft, H.; Griffiths, M.W. (1995) Specific oligonucleotide primers for detection of lecithinasepositive bacillus spp. by PCR. Appl. Environm. Microbiol. 61 (1), 98-102.
-9Seltmann, M.; Böhm, M.; Walter, T.; Schmitz, G.; Patterson, D.; Wieland, I. (1994) Isolation of
region specific single-copy probes from human chromosome 1q23  1q25. Cytogenet. Cell.
Genet. 67, 46-51.
Su, X.; Sullivan, T.F.; Bursztajn, S.; Berman, S.A. (1994) Optimized chemiluminescent detection
of DNA amplified in the exponential phase of PCR. BioTechn. 17 (4), 734-736.
Taveira, N.C.; Gomes, P.C.; Ferreira, M.O.S.; Pereira, J.M. (1994) Detection of HIV-1 DNA by
polymerase chain reaction incorporation of digoxigenin-11-dUTP and hybridization to immobilized
probes. Mol. Cell. Probes 8, 235-240.
Thomson, W.; Ollier, W. (1991) Non-radioactive ASO typing for class II -The way forward. Eur. J.
Immunogen. 19, 165-175.
Thümmler, F.; Beetz, A.; Rüdiger, W. (1990) Detection and partial sequence of a phytochrome
gene in the moss Ceratodon purpureus using the Polymerase Chain Reaction. FEBS 275 (1,2),
125-129.
Vilcek, S.; Herring, A. (1994) A single-tube method for the preparation of digoxigenin-labeled
probe from RNA by PCR. BioTechn. 16 (6), 1020.
Vlieger, A.M.; Medenblik, A.M.J.C.; van Gijlswijk, R.P.M.; Tanke, H.J., van der Ploeg, M.;
Gratama, J.W.; Raap, A.K. (1992) Quantitation of Polymerase Chain Reaction products by
hybridization-based assays with fluorescent, colorimetric, or chemiluminescent detection. Anal.
Biochem. 205, 1-7.
Woolford, A.J.; Dale, J.W. (1992) Simplified procedures for detection of amplified DNA using
fluorescent label incorporation and reverse probing. FEMS Microbiol. Let. 99, 311-316.
Yang, G.; Ulrich, P.P.; Aiyer, R.A.; Rawal, B.D.; Vyas, G.N. (1993) Detection of hepatitis B virus
in plasma using flow cytometric analyses of Polymerase Chain Reaction-amplified DNA
incorporation digoxigenin-11-dUTP. Blood 81 (4), 1083-1088.
Yun, Z.B.; Hornsleth, A. (1991) Production of digoxigenin-labeled parvovirus DNA probe by
PCR. Res. Virol. 142, 277-281.
Zakrzewska, K.; Ciappi, S.; Azzi, A. (1993) Polymerase chain reaction for synthesis of
digoxigenin-labelled DNA probe: application to parvovirus B19 and to polyomavirus BK. Mol.
Cell. Probes 7, 55-60.
B. RNA probes
Höltke, H.-J.; Kessler, C. (1990) Non-radioactive labeling of RNA transcripts in vitro with the
hapten digoxigenin (DIG); hybridization and ELISA-based detection. Nucleic Acids Res. 18 (19),
5843 - 5851.
Kietzmann, T.; Roth, U.; Freimann, S.; Jungermann, K. (1997) Arterial oxygen partial pressures
reduce the insulin-dependent induction of the perivenously located glukokinase in rat hepatocyte
cultures: mimicry of aterial oxygen pressures by H2O2. Biochem. J. 321, 17-20.
Rosemeyer, V.; Laubrock, A.; Seibl, R. (1995) Nonradioactive 3'-end-labeling of RNA molecules
of different lengths by terminal deoxynucleotidyltransferase. Anal. Biochem. 224, 446-449.
C. Oligonucleotide probes
a. 3'Endlabeling
Efremov, D.G.; Dimovski, A.J.; Efremov, G.D. (1991) Detection of ß-thalassemia mutations by
aso hybridization of PCR amplified DNA with digoxigenin ddUTP labeled oligonucleotides.
Hemoglobin 15 (6), 523-533.
- 10 Seidl, C.; Moritz, K.B. (1998) A novel UV-damaged DNA binding protein emerges during the
chromatin-eliminating cleavage period in Ascaris suum. Nucleic Acids Research Vol. 26 No. 3,
768-777.
Thomson, W.; Ollier, W. (1992) Non-radioactive aso typing for class II - The way forward. Eur. J.
Immunogen. 19, 169-175.
b. 3'Tailing
Mathieu, I.; Meyer, J.; Moulis, J.-M. (1992) Cloning, sequencing and expression in Escherichia
coli of the rubredoxin gene from Clostridium pasteurianum. Biochem. J. 285, 255-262.
Nickerson, D.A.; Kaiser, R.; Lappin, S.; Stewart, J.; Hood, L.; Landegren, U. (1990) Automated
DNA diagnostics using an ELISA-based oligonucleotide ligation assay. Proc. Natl. Acad. Sci. USA
87, 8923-8927.
Schmitz, G.G.; Walter, T.; Seibl, R.; Kessler, C. (1991) Non- radioactive labeling of
oligonucleotides in vitro with the hapten digoxigenin by tailing with Terminal Transferase. Anal.
Biochem. 192, 222-231.
Zuber, U.; Schumann, W. (1991) Short Communications: Tn5cos: a transposon for restriction
mapping of large plasmids using phage lambda terminase. Gene 103, 69-72.
c. Chemical synthesis (DIG-X-NHS ester)
Bürgi, W.; Keller, E.; Schmidheini, T. (1992) Nicht-enzymatische Markierung von Gen-Sonden
mit Digoxigenin für die Diagnose der Sichelzellmutation und der ß-Thalassämie. Schweiz. med.
Wschr. 122, 1567-1570.
Escarcellar, M.; Rodriguez-Frias, F.; Jardi, R.; San Segundo, B.; Eritja, R. (1992) Detection of
hepatitis B virus DNA in human serum. Samples: Use of digoxigenin-labeled
oligonucleotides as
modified primers for the Polymerase Chain Reaction. Anal. Biochem. 206,
36-42.
2. Detection techniques
A. NBT/BCIP detection
see e.g. "General papers"
Bosch, A., Gajardo, R., Diez, J.M., Pintó, R.M. (1996) Non isotopic automatable molecular
procedures for the detection of enteroviruses. Molecular and Cellular Probes, 10, 81 - 89.
Rihn, B.; Bottin, M.C.; Coulais, C.; Martinet, N. (1995) Evaluation of non-radioactive labeling and
detection of deoxyribonucleic acids. Part two: colorigenic methods and comparison with
chemiluminescent methods. J. Biochem. Biophys. Methods 30, 103-112.
B. Chemiluminescent detection
Allefs, J.J.H.M.; Salentijn, E.M.J.; Krens, F.A.; Rouwendal, G.J.A. (1990) Optimization of
non-radioactive southern blot hybridization: singly copy detection and reuse of blots. Nucleic Acids
Res. 18 (10), 3099-3100.
Didenko, V.V., Hornsby, P.J. (1996) Technical Note: A Quantitative Luminescence Assay
for Nonradioactive Nucleic Acid Probes. The Journal of Histochemistry and Cytochemistry,
44 (6), 657 - 660.
Dubitsky, A.; Brown, J.; Brandwein, H. (1992) Chemiluminescent detection of DNA on nylon
membranes. BioTechniques 13 (3), 392-399.
- 11 Girotti, S.; Ferri, E.; Ghini, S. (1991) Direct quantitative chemiluminescent assays for the
detection of viral DNA. Anal. Chimica Acta 255, 387-394.
Höltke, H.-J.; Sagner, G.; Kessler, C.; Schmitz, G. (1992) Sensitive chemiluminescent detection
of digoxigenin-labeled nucleic acids: A fast and simple protocol and its applications.
BioTechniques 12 (1), 104-113.
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5. Non-radioactive sequencing
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6. Non-radioactive RNase Protection Assay
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7. Non-radioactive Gel-Shift Assay
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10. Differential Display
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12. Miscellaneous
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