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Hidden Pitfalls of Using Onion Pollen in Molecular Research
Version 1
: Received: 21 December 2022 / Approved: 22 December 2022 / Online: 22 December 2022 (12:58:49 CET)
How to cite: Mardini, M.; Ermolaev, A.; Khrustaleva, L. Hidden Pitfalls of Using Onion Pollen in Molecular Research. Preprints 2022, 2022120429. https://doi.org/10.20944/preprints202212.0429.v1 Mardini, M.; Ermolaev, A.; Khrustaleva, L. Hidden Pitfalls of Using Onion Pollen in Molecular Research. Preprints 2022, 2022120429. https://doi.org/10.20944/preprints202212.0429.v1
Abstract
There is little information on the use of pollen in molecular research, despite the increased interest to genome editing by pollen mediated transformation. PCR is useful tool as an express-method to evaluate editing results before pollination. A direct PCR protocol for pollen suspension has been adapted without the need for DNA pre-extraction. We showed that pollenkitt is a limiting factor for successful PCR on pollen. A simple pre-washing step of pollen suspension was able to eliminate the pollenkitt and enormously affect the PCR results. All currently existing methods of delivery of the editing system to pollen are carried out in a wet medium. Our pollenkitt study helped us develop a simple and effective pollination method using wetted onion pollen grains.
Keywords
onion; PCR inhibition; pollenkitt; wetted pollen; pollination
Subject
Biology and Life Sciences, Biochemistry and Molecular Biology
Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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