Do NTP's precipitate with ethanol? How efficiently?
Anton Scott Goustin
asg at cmb.biosci.wayne.edu
Thu Feb 15 15:21:44 EST 1996
The information in Chi-Kuang's reply (above) is quite misleading.
Deoxyribonucleotide triphosphates are quite insoluble in mixtures of
EtOH and NaCl. For example, if you precipitate a randon-primer reaction
by first raising the NaCl to 0.3 M and then adding 2.5 volumes of EtOH,
a substantial fraction of your dNTPs will come down with the DNA. The
solubility of dNTPs, however, is substantially reduced in ammonium
salts. For example, a clean separation between dNTPs and DNA can be
achieved by first raising the concentration of ammonium chloride to 4
molar and then adding 2.5 vol EtOH.
Ppttn of dNTPs also depends on the base. G is most insoluble, A is
most soluble. The ribo versions of each NTP are also more soluble than
the deoxy versions.
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