Use of FTA® classic cards for epigenetic analysis of sperm DNA | Zendy

Olga Serra | Zendy; Raffaele Frazzi | Zendy; Alessio Perotti | Zendy; Lorenzo Barusi | Zendy; Annamaria Buschini | Zendy

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Use of FTA® classic cards for epigenetic analysis of sperm DNA

Author(s) -

Olga Serra,

Raffaele Frazzi,

Alessio Perotti,

Lorenzo Barusi,

Annamaria Buschini

Publication year - 2018

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/btn-2017-0101

Subject(s) - epigenetics , dna , bisulfite , elution , dna methylation , biology , sperm , dna extraction , bisulfite sequencing , str analysis , quantitative analysis (chemistry) , chromatography , computational biology , microbiology and biotechnology , genetics , chemistry , polymerase chain reaction , microsatellite , gene , gene expression , allele

FTA® technologies provide the most reliable method for DNA extraction. Although FTA technologies have been widely used for genetic analysis, there is no literature on their use for epigenetic analysis yet. We present for the first time, a simple method for quantitative methylation assessment based on sperm cells stored on Whatman FTA classic cards. Specifically, elution of seminal DNA from FTA classic cards was successfully tested with an elution buffer and an incubation step in a thermocycler. The eluted DNA was bisulfite converted, amplified by PCR, and a region of interest was pyrosequenced.

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