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We processed one negative extraction control for each six fossil samples extracted (total of 28 blank extraction controls) and followed these samples through PCR amplification.
For each set of extractions a blank extraction was performed using all the reagents minus the DNA extract (this was performed at least once for every 10 insects).
A blank extraction was also performed in parallel to the mosasaur DNA extraction.
Negative controls were run using blank extraction samples for each PCR run.
They corresponded to one extraction blank (extraction without sample tissue), one PCR blank (a tube with PCR mix without DNA) and one aerosol blank (a PCR blank with the cap open during the whole manipulation).
Blank extraction and PCR controls were used in each reaction and only DNA sequences which could be replicated from at least two independent amplifications of each primer pair were used in the subsequent analyses.
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The sample was extracted with blank extractions to monitor against contamination.
Blank extractions were included every five samples to monitor for contamination.
Blank extractions were included in each batch to monitor for cross-over contamination.
DNA and blank extractions from both methods, MoBio and SO, were purified and concentrated using Wizard prep SV columns (Promega).
Blank extractions were performed with every batch.
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