Improved library preparation protocols for amplicon sequencing-based noninvasive fetal genotyping for RHD-positive D antigen-negative alleles

DOI PubMed 研究データあり 被引用文献1件 参考文献1件 オープンアクセス

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<jats:title>Abstract</jats:title><jats:sec> <jats:title>Objective</jats:title> <jats:p>We aimed to simplify our fetal <jats:italic>RHD</jats:italic> genotyping protocol by changing the method to attach Illumina’s sequencing adaptors to PCR products from the ligation-based method to a PCR-based method, and to improve its reliability and robustness by introducing unique molecular indexes, which allow us to count the numbers of DNA fragments used as PCR templates and to minimize the effects of PCR and sequencing errors.</jats:p> </jats:sec><jats:sec> <jats:title>Results</jats:title> <jats:p>Both of the newly established protocols reduced time and cost compared with our conventional protocol. Removal of PCR duplicates using UMIs reduced the frequencies of erroneously mapped sequences reads likely generated by PCR and sequencing errors. The modified protocols will help us facilitate implementing fetal <jats:italic>RHD</jats:italic> genotyping for East Asian populations into clinical practice.</jats:p> </jats:sec>

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  • BMC Research Notes

    BMC Research Notes 14 2021-09-26

    Springer Science and Business Media LLC

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