Enabling the Spaceflight Methylome: DNA Isolated from Plant Tissues Preserved in RNAlater® Is Suitable for Bisulfite PCR Assay of Genome Methylation

Collin E. LeFrois, Mingqi Zhou, David Moraga Amador, Natasha Sng, Anna-Lisa Paul, Robert J. Ferl
2020 Gravitational and space research : publication of the American Society for Gravitational and Space Research  
AbstractSpaceflight has a unique set of abiotic conditions to which plants respond by orchestrating genome-wide alterations to their transcriptome. The methods for preserving plants for RNA analysis are well-established and proven over multiple missions, but, methods for investigating the possible epigenetic mechanisms that may contribute to the transcriptome alteration are not well-developed for the confining limitations of the International Space Station (ISS). Currently, the methods used to
more » ... solate genomic DNA and to perform epigenetic analyses are ideal for frozen plants, as opposed to plants stored in RNAlater®—a high salt solution that chemically suspends all cellular activity and is typically used on the ISS. Therefore, we developed a method for extracting high-quality genomic DNA suitable for epigenetic analysis from Arabidopsis thaliana (Arabidopsis) plants that were preserved with the current preservation system aboard the ISS—fixation in RNAlater® using Kennedy Space Center Fixation Tubes (KFTs).
doi:10.2478/gsr-2016-0010 fatcat:7ncupb7qa5ckjkeucehgvnag5m