<abstract xmlns="http://www.w3.org/1999/xhtml">Because of difficulties during the fixation in space and the often reported enhanced expression of stress-related genes in space experiments, we investigated the possible effect of fixation on gene expression. Comparing two fixatives (RNAlater® and 70% ethanol), two-day-old <italic>Brassica rapa</italic> seedlings were either fixed by gradual exposure or immediate and complete immersion in fixative for two days. Neither fixative yielded high amounts of rRNA; RNAlater® resulted in higher RNA yield in shoot tissue but qPCR data showed higher yield in ethanol-fixed material. qPCR analyses showed strongly enhanced transcripts of stress-related genes, especially in RNAlater®-fixed material. The data suggest that fixation artefacts may be partially responsible for effects commonly attributed to space syndromes.</abstract>

Because of difficulties during the fixation in space and the often reported enhanced expression of stress-related genes in space experiments, we investigated the possible effect of fixation on gene expression. Comparing two fixatives (RNAlater® and 70% ethanol), two-day-old Brassica rapa seedlings were either fixed by gradual exposure or immediate and complete immersion in fixative for two days. Neither fixative yielded high amounts of rRNA; RNAlater® resulted in higher RNA yield in shoot tissue but qPCR data showed higher yield in ethanol-fixed material. qPCR analyses showed strongly enhanced transcripts of stress-related genes, especially in RNAlater®-fixed material. The data suggest that fixation artefacts may be partially responsible for effects commonly attributed to space syndromes.

Beware of Fixation—It Might Affect Your Experiments

National Institute of Crop Science (NICS)

Gravitational and Space Research

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

{"article-title":"Beware of Fixation\u2014It Might Affect Your Experiments"}

Because of difficulties during the fixation in space and the often reported enhanced expression of stress-related genes in space experiments, we investigated...

Beware of Fixation—It Might Affect Your Experiments

Article Category: Research Article

Pubblicato online: 18 lug 2020

Pagine: 47 - 57

DOI: https://doi.org/10.2478/gsr-2016-0012

Parole chiave
Ethanol, Fixation, Gene Expression, Spaceflight, RNAlater®

© 2020 Myoung-Ryoul Park et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Beware of Fixation—It Might Affect Your Experiments

Article Category: Research Article

Pubblicato online: 18 lug 2020

Pagine: 47 - 57

DOI: https://doi.org/10.2478/gsr-2016-0012

Parole chiaveEthanol, Fixation, Gene Expression, Spaceflight, RNAlater®

© 2020 Myoung-Ryoul Park et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Parole chiave
Ethanol, Fixation, Gene Expression, Spaceflight, RNAlater®