title: Extracellular embryo genomic DNA and its potential for genotyping applications.
abstract: BACKGROUND
  Preimplantation genetic diagnosis (PGD) currently relies on biopsy of one or few embryo cells. Our aim was to evaluate the embryo extracellular matrices (spent medium and blastocoele fluid) as source of DNA for embryo genotyping.
  We first evaluated the amplifiability and the amount of genomic DNA in spent embryo culture media from day 3 (n = 32) and day 5/6 (n = 54). Secondly, we evaluated the possibility to genotype theMTHFRpolymorphism C677T from media at day 5/6 (n = 8) and blastocoele fluids (n = 9) by direct sequencing. The C677T polymorphism detection rate was 62.5 and 44.4% in medium and fluid, respectively.
  
  
  CONCLUSIONS
  A noninvasive approach for embryo genotyping was possible, but still with limitations due to low detection rate and possible allele dropout.
authors.affiliation: Department of Biomolecular Sciences, University of Urbino 'Carlo Bo', 61029 Urbino (PU), Italy; Department of Biomolecular Sciences, University of Urbino 'Carlo Bo', 61029 Urbino (PU), Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Galluzzi
authors.affiliation: IVF Unit, 'Cervesi' Hospital Cattolica, 47841 Cattolica (RN), Italy; IVF Unit, 'Cervesi' Hospital Cattolica, 47841 Cattolica (RN), Italy.
authors.forename: Simone
authors.initials: S
authors.lastname: Palini
authors.affiliation: IVF Unit, 'Cervesi' Hospital Cattolica, 47841 Cattolica (RN), Italy; IVF Unit, 'Cervesi' Hospital Cattolica, 47841 Cattolica (RN), Italy.
authors.forename: Silvia De
authors.initials: S
authors.lastname: Stefani
authors.affiliation: Department of Biomolecular Sciences, University of Urbino 'Carlo Bo', 61029 Urbino (PU), Italy; Department of Biomolecular Sciences, University of Urbino 'Carlo Bo', 61029 Urbino (PU), Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Andreoni
authors.affiliation: IVF Unit, 'Cervesi' Hospital Cattolica, 47841 Cattolica (RN), Italy; IVF Unit, 'Cervesi' Hospital Cattolica, 47841 Cattolica (RN), Italy.
authors.forename: Mariangela
authors.initials: M
authors.lastname: Primiterra
authors.affiliation: Department of Biomolecular Sciences, University of Urbino 'Carlo Bo', 61029 Urbino (PU), Italy; Department of Biomolecular Sciences, University of Urbino 'Carlo Bo', 61029 Urbino (PU), Italy.
authors.forename: Aurora
authors.initials: A
authors.lastname: Diotallevi
authors.affiliation: IVF Unit, 'Cervesi' Hospital Cattolica, 47841 Cattolica (RN), Italy; IVF Unit, 'Cervesi' Hospital Cattolica, 47841 Cattolica (RN), Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Bulletti
authors.affiliation: Department of Biomolecular Sciences, University of Urbino 'Carlo Bo', 61029 Urbino (PU), Italy; Department of Biomolecular Sciences, University of Urbino 'Carlo Bo', 61029 Urbino (PU), Italy.
authors.forename: Mauro
authors.initials: M
authors.lastname: Magnani
keywords: MTHFR; blastocoele fluid; culture medium; gDNA; human embryos