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Optimization of Chelex 100 resin-based extraction of genomic DNA from dried blood spots by Neta Simon; Jaclyn Shallat; Corey Williams Wietzikoski; Whitney E Harrington is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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## Abstract Dried blood spots (DBS) are widely utilized as part of universal newborn screening and as a means of transporting samples from field sites. We use DBS from African field sites to assess for rare maternal-fetal cell exchange during pregnancy known as microchimerism. We aimed to develop a protocol to maximize the quantity of high-quality genomic DNA (gDNA) extracted from DBS. The total gDNA yield obtained from control DBS utilizing a Qiagen-based protocol and a Chelex^®^ 100 resin-based protocol was first compared. Variations of the Chelex^®^ protocol were subsequently tested to develop an optimized protocol. The gDNA was quantified by qPCR targeting the human beta-globin gene. DNA yield for a given experimental condition was normalized to a Chelex^®^ control performed on the same day, and the total yields were compared using a Student’s __t__-test. The control Chelex^®^ protocol yielded 590% more DNA than the QIAamp^®^ DNA Blood Mini Kit . The absolute efficiency of the control Chelex^®^ protocol was 54%, compared to an absolute efficiency of 9% for the QIAamp^®^ DNA Blood Mini Kit. Modification of the Chelex^®^ protocol to include a second heat precipitation from the sa

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Author

Neta Simon; Jaclyn Shallat; Corey Williams Wietzikoski; Whitney E Harrington

Published

2020

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)