damage for organellar DNA
نویسندگان
چکیده
Our objective was to use long PCR to determine the copy number of ptDNA and mtDNA in total DNA extracted from tissue samples. In previous work, the efficiency of amplification of DNA decreased as the target sequence length increased above about 1 kb, so that DNA quantification using long PCR was considered unreliable (Arezi et al. 2003). Here, we used a long-PCR endpoint procedure to amplify organellar DNA fragments of ~11 kb and performed quantification using gel electrophoresis, ethidium bromide (EtBr)-DNA fluorescence, and DNA mass standards. We achieved an efficiency of ~100% and validated the method for assessing orgDNA damage. These ptDNA and mtDNA primers and long-PCR conditions were used:
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