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Covalent Attachment of DNA to Agarose: Improved Synthesis and Use in Affinity Chromatography
Open accessAbstract
DNA has been covalently linked to insoluble matrices of agarose (Sepharose) in high yield using cyanogen bromide activation. Both double‐stranded and single‐stranded DNA have been coupled with yields up to 225 nmol/mg dry weight Sepharose or 3–8 μmol nucleotide phosphate/ml bed volume. The DNA‐Sepharose has been used for (a) the affinity chromatography of various enzymes (Escherichia coli DNA polymerase I and RNA polymerase) from crude extracts or after initial purification steps, resulting in high yields and degrees of purification, and for (b) nucleic acid hybridization. The DNA‐Sepharose is stable to high temperature, prolonged storage, and in the case of single‐stranded DNA, can be washed with NaOH to destroy nuclease activity and to release any digested oligonucleotides or mononucleotides.
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Cited by
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DNA N-glycosidases: properties of uracil-DNA glycosidase from Escherichia coli.
Lindahl, T., Ljungquist, S., Siegert, W., Nyberg, B., & Sperens, B. (1977) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)40386-3 · Cited by 522
Enzymatic in vitro synthesis of globin genes
Efstratiadis, A., Kafatos, F. C., Maxam, A. M., & Maniatis, T. (1976) · Cell · doi:10.1016/0092-8674(76)90027-1 · Cited by 342
E. coli and the etiology of human PBC: Antimitochondrial antibodies and spreading determinants
Yang, Y., Choi, J., Chen, Y., Invernizzi, P., Yang, G., Zhang, W., Shao, T., Jordan, F., Nemeria, N. S., Coppel, R. L., et al. (2022) · Hepatology · doi:10.1002/hep.32172 · Cited by 44
Metadata
- Authors
- ARNDT‐JOVIN, D. J., JOVIN, T. M., BÄHR, W., MARQUARDT, M., & FRISCHAUF, A.
- Venue
- European Journal of Biochemistry
- Publisher
- Wiley
- Published
- 1975-06
- Volume
- 54
- Issue
- 2
- Pages
- 411-418
- ISSN
- 0014-2956 (ISSN-L)
- 1432-1033
- OpenAlex
- W1526634010
- Cited by
- 206
- References
- 0
- FWCI
- 5.71
- Type
- journal article
- Language
- en
- Open access
- bronze
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- The paper
- ARNDT‐JOVIN, D. J., JOVIN, T. M., BÄHR, W., MARQUARDT, M., & FRISCHAUF, A. (1975). Covalent Attachment of DNA to Agarose: Improved Synthesis and Use in Affinity Chromatography. European Journal of Biochemistry. https://doi.org/10.1111/j.1432-1033.1975.tb04151.x
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
