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When tick larvae were found on squirrel carcasses, DNA extractions were carried out on a pooled sample of up to 10 larvae from each squirrel.
For carcass analysis, lipid extraction was performed as described [ 13] and body fat percentage was determined.
These DXA measures were, however, validated previously in piglets using biochemical assessment of carcass N content and lipid extraction to determine lean and fat mass, respectively.
However, material for this study was also collected from wild boar carcasses, allowing only viral DNA extraction.
Urine was punctured from the urinary bladder immediately after extraction of the organs from the carcass.
After removing the digestive tract, the remaining tissue (i.e., the fat body, reduced reproductive organs, exoskeleton, trachea, and muscles) was separated from the thorax and this abdominal carcass was preserved at -80°C for subsequent RNA extraction.
To resolve this issue, we extracted genomic DNA from 2940 muscle samples taken from the phenotyped carcasses by proteinase K digestion followed by phenol chloroform isoamyl alcohol extraction and ethanol precipitation (Sambrook et al. 1989).
Glands and the remaining body carcass were stored separately in RNAlater® (Ambion) for subsequent RNA extraction.
Carcasses of the dissected mosquitoes were stored at −20 °C until DNA extraction.
We therefore conclude that the re-extraction of DNA from tissue samples taken from the carcass at slaughter effectively solved the misidentification problem.
Extraction efficiency of PBDE-47 was 90 95% in blank carcasses fortified with PBDE-47 at concentrations encompassing observed levels in experimental fish.
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