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In vitro incorporation of tritium into native DNA
Author(s) -
DopplerBernardi Françoise,
Felsenfeld Gary
Publication year - 1969
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.1969.360080604
Subject(s) - tritium , chemistry , dna , in vitro , in vivo , denaturation (fissile materials) , nucleotide , duplex (building) , specific activity , biochemistry , molecule , radiochemistry , nuclear chemistry , organic chemistry , enzyme , physics , gene , nuclear physics , biology , microbiology and biotechnology
An exchange method is described for producing tritium‐labeled native DNA in vitro with minimal physical damage to the DNA. Tritium‐labeled calf thymus DNA prepared in this way has a specific activity of about 100 μCi/mmole of nucleotide (i.e., about 2 × 10 8 dpm/mmole). Sedimentation velocity at neutral and alkaline pH indicate that the product has an average of two single strand breaks per duplex molecule of molecular weight 6 × 10 6 daltons. The optical and thermal denaturation properties of the product are those of native DNA. The method should be particularly useful for labeling DNA from organisms that cannot be labeled conveniently in vivo .