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The mitochondrial genomic DNA was extracted from buccal mucosal cells by using buccal cell DNA extraction kit, BuccalQuick™ and specially designed foam swab, Easy-Swab™ (TrimGen, Maryland, USA).
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However, the lack of consistent statistically significant enrichment for the smoker phenotype with GSEA analysis taken with the degradation in RNA derived from buccal cells highlight the difficulties to be expected when using buccal-cell RNA for differential expression testing.
Cells for DNA-preparation were collected directly by the patients using buccal swabs.
Biological material was collected using buccal swabs.
Buccal cell DNA extractions were performed using Puregene Buccal Cell Core Kits (Qiagen) according to the manufacturer's instructions, except all centrifugations were conducted at 4°C instead of room temperature.
Buccal cell and CD4+ lymphocyte DNA extractions were performed using Puregene buccal cell core kits (Qiagen Sciences, Germantown, Maryland, United States) according to the manufacturer's instructions, except all centrifugations were conducted at 4°C instead of room temperature.
DNA was extracted by using the Gentra Puregene Buccal Cell Kit (QIAGEN AB, Stockholm, Sweden) and dissolved in a 100-μL DNA hydration solution (provided with the kit).
For the buccal rinse, the buccal cells were pelleted and DNA was extracted using the Puregene Buccal Cell Kit following manufacturer's instructions (Qiagen Sciences, Valencia, CA).
Total DNA was then extracted using the Qiagen buccal cell spin protocol (Qiagen Ltd., Crawley, UK).
Genotyping has become more cost-effective and less invasive with the use of buccal cell sampling.
The cells were collected by centrifugation at 1200 g and DNA was isolated using the Gentra Puregene buccal cell kit (Qiagen, Valencia, CA, USA).
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