3 ml of an EDTA‐preserved whole blood sample of de novo probands’ healthy parents was used for genomic DNA purification, using the Gentra Puregene Blood Kit (Quiagen, Germany) in accordance with the manufacturer's protocol, and stored at −80°C.
In order to affirm the absence of an OI‐causative pathogenic variant in healthy parents, Sanger sequencing of an exon carrying a subject's pathogenic variant was performed. PCR amplification, Sanger sequencing, and analysis of the sequencing products were each performed as described in previous studies (Ho Duy et al.,2016 ; Zhytnik et al., 2017 ).
Sequence products were also aligned to the GenBank human reference genome sequences of COL1A1 (gDNA NG_007400.1, complementary (cDNA) NM_000088.3) and COL1A2 (gDNA NG_007405.1, cDNA NM_000089.3). The datasets used and analyzed during the study are available from the corresponding author upon reasonable request.
In order to affirm the absence of an OI‐causative pathogenic variant in healthy parents, Sanger sequencing of an exon carrying a subject's pathogenic variant was performed. PCR amplification, Sanger sequencing, and analysis of the sequencing products were each performed as described in previous studies (Ho Duy et al.,
Sequence products were also aligned to the GenBank human reference genome sequences of COL1A1 (gDNA NG_007400.1, complementary (cDNA) NM_000088.3) and COL1A2 (gDNA NG_007405.1, cDNA NM_000089.3). The datasets used and analyzed during the study are available from the corresponding author upon reasonable request.