In vitro incorporation of tritium into native DNA

✍ Scribed by Françoise Doppler-Bernardi; Gary Felsenfeld

• Book ID: 102762293
• Publisher: Wiley (John Wiley & Sons)
• Year: 1969
• Tongue: English
• Weight: 540 KB
• Volume: 8
• Category: Article
• ISSN: 0006-3525
• DOI: 10.1002/bip.1969.360080604

✦ Synopsis

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 pCi/mmole of nucleotide (i.e., about 2 x 108 dpm/mmole). Sedimentation velocity a t neutral and alkaline pH indicate that the product has an average of two single strand breaks per duplex molecule of molecular weight 6 X 106 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.