High-Molecular-Weight Genomic DNA Extraction from Recalcitrant Australian Plants: An Optimised CTAB Protocol for Anigozanthos
- 1The University of Western Australia
Abstract
High-phenolic plant genera present a major technical limitation in genomic research. Standard extraction approaches that perform reliably across diverse flora often perform poorly when applied to recalcitrant taxa, producing low DNA yield and integrity incompatible with sequencing requirements. The genus Anigozanthos (Kangaroo paws) from the family Haemodoraceae exemplifies this problem. We identified key physicochemical factors governing extraction failure in this genus and resolved them through targeted modifications to lysis chemistry and contaminant management. The resulting protocol achieved a near threefold improvement in DNA purity, substantially reducing contaminant carry over and consistently yielded high-integrity, long DNA fragments (DIN > 7) across a diverse sample set spanning cultivated and wild material across four diverse genera of Haemodoraceae. We also tested a straightforward purity assessment framework that can be implemented in any standard molecular laboratory, enabling rapid pre-submission quality assessment without the need for specialised equipment. Together these advances open a practical path to genomic characterisation of Anigozanthos that establishes a transferable model for genomic research across Australia ' s chemically complex native flora.
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BibTeX
@misc{rajput2026highmolecularweight,
title = {{High-Molecular-Weight Genomic DNA Extraction from Recalcitrant Australian Plants: An Optimised CTAB Protocol for Anigozanthos}},
author = {R. Rajput and L. Saha and Z. Ahmed and P. Naiker and L. Do and A. Bisset and C. Hooper},
howpublished = {bioRxiv},
year = {2026},
month = aug,
doi = {10.64898/2026.08.29.741951},
url = {https://www.biorxiv.org/content/10.64898/2026.08.29.741951v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - High-Molecular-Weight Genomic DNA Extraction from Recalcitrant Australian Plants: An Optimised CTAB Protocol for Anigozanthos
AU - R. Rajput
AU - L. Saha
AU - Z. Ahmed
AU - P. Naiker
AU - L. Do
AU - A. Bisset
AU - C. Hooper
PY - 2026
DA - 2026/08/29
PB - bioRxiv
DO - 10.64898/2026.08.29.741951
UR - https://www.biorxiv.org/content/10.64898/2026.08.29.741951v1
AB - High-phenolic plant genera present a major technical limitation in genomic research. Standard extraction approaches that perform reliably across diverse flora often perform poorly when applied to recalcitrant taxa, producing low DNA yield and integrity incompatible with sequencing requirements. The genus Anigozanthos (Kangaroo paws) from the family Haemodoraceae exemplifies this problem. We identified key physicochemical factors governing extraction failure in this genus and resolved them through targeted modifications to lysis chemistry and contaminant management. The resulting protocol achieved a near threefold improvement in DNA purity, substantially reducing contaminant carry over and consistently yielded high-integrity, long DNA fragments (DIN > 7) across a diverse sample set spanning cultivated and wild material across four diverse genera of Haemodoraceae. We also tested a straightforward purity assessment framework that can be implemented in any standard molecular laboratory, enabling rapid pre-submission quality assessment without the need for specialised equipment. Together these advances open a practical path to genomic characterisation of Anigozanthos that establishes a transferable model for genomic research across Australia ' s chemically complex native flora.
ER -