ABSTRACT
Some primers frequently used in the double-stranded dideoxy DNA sequencing technique with radioactive markers are not suited for fluorescent detection. In fact oligonucleotides have a different annealing efficiency related to their base sequence, and this is reflected in nonequivalent results, particularly in fluorescent automated DNA sequencing. We present a method for the evaluation of primer performance in automated DNA sequencers and show its application to the search for a better set of primers for pBluescript vector.
Subject(s)
Base Sequence , DNA/genetics , Genetic Techniques , DNA Probes , Evaluation Studies as Topic , Molecular Probe Techniques , Molecular Sequence Data , ThermodynamicsABSTRACT
The apparent Km for cytochrome c of cytochrome oxidase does not change but the Vmax decreases in synaptosomes and non-synaptic mitochondria isolated from the cerebral cortex as a whole of 30-month-old rats compared with 4-month-old ones. When the subcellular organelles are submitted to stressful conditions, namely incubation in media of altered osmolality, the percentage of cytochrome oxidase activity released is much higher in senescent rats. The activity of cytochrome oxidase evaluated in non-synaptic mitochondria and synaptosomes isolated from cortical and subcortical regions and cerebellum of rats aged 4 and 30 months shows a highly significant decrease (P less than 0.001) in the parietotemporal cortex of senescent rats (both in non-synaptic mitochondria and synaptosomes) and in the cerebellum (in synaptosomes).
