Allele-specific mRNA abundances were measured using PyrosequencerAB (Biotage AB, now Qiagen) following manufacturer's instructions for sample preparation and pyrosequencing reactions.
Abiding by service provider's instructions for sample collection and shipping, genotyping was conducted via quantitative polymerase chain reaction (q-PCR -based technologies (Transnetyx, Inc., Cordova, TN).
Total RNA extraction was performed by homogenizing the fat tissue samples with TRIZOL® (TRI reagent, Invitrogen, Carlsbad, CA, USA) following the manufacturer's instructions for samples with high fat content.
Subjects who selected a stool test were provided the Hb/Hp Complex ELISA stool collection kit, instructions for sample collection, and were requested to provide the fecal sample to the testing lab.
Reptile owners who agreed to participate in the study received detailed written sampling instructions, materials for sampling (small plastic container, plastic spoon, and Aimes agar gel medium transport swabs (Copan Italia S.P.A, Brescia, Italy) by mail one to two weeks before sampling.
Instructions for CSNT Sampling; Cornell Nutrient Management Spear Program: http://nmsp.cals.cornell.edu/projects/nitrogenforcorn/StalkNtest.pdf.edu/projects/nitrogenforcorn/StalkNtest.pdf
Amplified RNA from tumor or cell samples was handled according to the manufacture's instructions for eukaryotic sample processing and hybridized to the arrays.
In addition, the investigators observed that the instructions for pharyngeal samples were less well adhered to than the instructions for nasal samples.
