AI Article Synopsis
- - We simplified and improved the fetal RHD genotyping protocol by switching from a ligation-based method to a PCR-based method for attaching Illumina's sequencing adaptors, making the process more efficient.
- - The new approach includes unique molecular indexes (UMIs) that help count DNA fragments and reduce errors from PCR and sequencing, enhancing the reliability of the results.
- - Overall, the updated protocols save time and money compared to the old method and are designed to make fetal RHD genotyping easier to implement in East Asian clinical settings.
Article Abstract
Objective: We aimed to simplify our fetal RHD 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.
Results: 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 RHD genotyping for East Asian populations into clinical practice.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8474863 | PMC |
| http://dx.doi.org/10.1186/s13104-021-05793-4 | DOI Listing |
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