Assessing the Performance of Dried-Blood-Spot DNA Extraction Methods in Next Generation Sequencing

International Journal of Neonatal Screening
Miyono HendrixSuzanne K Cordovado

Abstract

An increasing number of newborn screening laboratories in the United States and abroad are moving towards incorporating next-generation sequencing technology, or NGS, into routine screening, particularly for cystic fibrosis. As more programs utilize this technology for both cystic fibrosis and beyond, it is critical to identify appropriate DNA extraction methods that can be used with dried blood spots that will result in consistent, high-quality sequencing results. To provide comprehensive quality assurance and technical assistance to newborn screening laboratories wishing to incorporate NGS assays, CDC's Newborn Screening and Molecular Biology Branch designed a study to evaluate the performance of nine commercial or laboratory-developed DNA extraction methods that range from a highly purified column extraction to a crude detergent-based no-wash boil prep. The DNA from these nine methods was used in two NGS library preparations that interrogate the CFTR gene. All DNA extraction methods including the cruder preps performed reasonably well with both library preps. One lower-concentration, older sample was excluded from one of the assay evaluations due to poor performance across all DNA extraction methods. When 84 samples, versus ...Continue Reading

References

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Oct 20, 2020·International Journal of Neonatal Screening·Cristina Gutierrez-MateoRongcong Wu

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Citations

Jun 15, 2021·Frontiers in Genetics·Ziga I RemecMarusa Debeljak
Aug 14, 2021·Molecular Genetics and Metabolism·Edward R B McCabe

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Methods Mentioned

BETA
blood collection
PCR
Assay
exome sequencing

Software Mentioned

GATK
Picard SAMtoBAM
MiSeqReporter
FreeBayes
MiSeq Reporter
AddOrReplaceReadGroups
NSQAP
SeqScape
vcflib suite
Picard

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