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Metal chelate affinity precipitation of RNA and purification of plasmid DNA

Balan, S; Murphy, J; Galaev, Igor LU ; Kumar, Ashok LU ; Fox, G; Mattiasson, Bo LU and Willson, R (2003) In Biotechnology Letters 25(13). p.1111-1116
Abstract
The affinity of metal chelates for amino acids, such as histidine, is widely used in purifying proteins, most notably through six-histidine `tails'. We have found that metal affinity interactions can also be applied to separation of single-stranded nucleic acids through interactions involving exposed purines. Here we describe a metal affinity precipitation method to resolve RNA from linear and plasmid DNA. A copper-charged copolymer of N-isopropyl acrylamide (NIPAM) and vinyl imidazole (VI) is used to purify plasmid from an alkaline lysate of E. coli. The NIPAM units confer reversible solubility on the copolymer while the imidazole chelates metal ions in a manner accessible to interaction with soluble ligands. RNA was separated from the... (More)
The affinity of metal chelates for amino acids, such as histidine, is widely used in purifying proteins, most notably through six-histidine `tails'. We have found that metal affinity interactions can also be applied to separation of single-stranded nucleic acids through interactions involving exposed purines. Here we describe a metal affinity precipitation method to resolve RNA from linear and plasmid DNA. A copper-charged copolymer of N-isopropyl acrylamide (NIPAM) and vinyl imidazole (VI) is used to purify plasmid from an alkaline lysate of E. coli. The NIPAM units confer reversible solubility on the copolymer while the imidazole chelates metal ions in a manner accessible to interaction with soluble ligands. RNA was separated from the plasmid by precipitation along with the polymer in the presence of 800 mM NaCl. Bound RNA could be recovered by elution with imidazole and separated from copolymer by a second precipitation step. RNA binding showed a strong dependence on temperature and on the type of buffer used. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biotechnology Letters
volume
25
issue
13
pages
1111 - 1116
publisher
Springer
external identifiers
  • pmid:12889823
  • wos:000183441800018
  • scopus:0041360009
ISSN
1573-6776
DOI
10.1023/A:1024148316322
language
English
LU publication?
yes
id
72cf6d0a-c6c7-4799-85c2-6098942707e7 (old id 129078)
date added to LUP
2007-06-29 16:05:16
date last changed
2017-01-01 07:11:30
@article{72cf6d0a-c6c7-4799-85c2-6098942707e7,
  abstract     = {The affinity of metal chelates for amino acids, such as histidine, is widely used in purifying proteins, most notably through six-histidine `tails'. We have found that metal affinity interactions can also be applied to separation of single-stranded nucleic acids through interactions involving exposed purines. Here we describe a metal affinity precipitation method to resolve RNA from linear and plasmid DNA. A copper-charged copolymer of N-isopropyl acrylamide (NIPAM) and vinyl imidazole (VI) is used to purify plasmid from an alkaline lysate of E. coli. The NIPAM units confer reversible solubility on the copolymer while the imidazole chelates metal ions in a manner accessible to interaction with soluble ligands. RNA was separated from the plasmid by precipitation along with the polymer in the presence of 800 mM NaCl. Bound RNA could be recovered by elution with imidazole and separated from copolymer by a second precipitation step. RNA binding showed a strong dependence on temperature and on the type of buffer used.},
  author       = {Balan, S and Murphy, J and Galaev, Igor and Kumar, Ashok and Fox, G and Mattiasson, Bo and Willson, R},
  issn         = {1573-6776},
  language     = {eng},
  number       = {13},
  pages        = {1111--1116},
  publisher    = {Springer},
  series       = {Biotechnology Letters},
  title        = {Metal chelate affinity precipitation of RNA and purification of plasmid DNA},
  url          = {http://dx.doi.org/10.1023/A:1024148316322},
  volume       = {25},
  year         = {2003},
}