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Some samples with low yields of NA PCR amplicons (see below) were reextracted from fecal samples by organic extraction with TRIzol (Life Technologies) using 900 μl reagent per 100 μl sample.
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The GA3 was reextracted from the residue as mentioned above.
When PCR failed, DNA was reextracted from original cultures, and genotyping was attempted again.
We previously analyzed the dust samples from the first round of dust collection for nine PAHs (Whitehead et al. 2009); however, for consistency, the dust samples from the first round of dust collection were reextracted and reanalyzed alongside the samples from the second round of dust collection according to the protocol described below.
Supernatants were reextracted with 2 ml of 70% ethanol.
After centrifugation (16000 g, 5 min, 4°C), supernatants were kept in clean tubes whereas pellets were reextracted with 1 ml 5% HClO4.
The organic phases were reextracted with water.
Samples failing this test were reextracted.
The resulting mixtures were filtered and the residues were reextracted in the same conditions.
The residues were reextracted again with chloroform/methanol mixture for 2 hr and evaporated under vacuum condition.
The extract was filtered, and the plant leaves were reextracted twice more according to the same method.
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