A modified CTAB method for high-molecular-weight DNA preparation from deep-sea holothurians

ABSTRACTWith the features as global distribution and all depths dwelling, holothurians function an ideal model for the study of deep-sea adaptation. Acquisition of high-molecular-weight DNA is the basic requirement for molecular biology analyses. However, extraction of high-molecular-weight DNA from deep-sea holothurians is difficult by degradation and contamination. Here, we presented a modified CTAB (lysis buffer with cetyl trimethyl ammonium bromide) method and compared its extraction efficiency for deep-sea holothurians with other five common methods, i.e. the Tiangen Biotech kit, Qiagen kit, Omega Bio-tek kit, Sangon Biotech kit, and SDS (lysis buffer with sodium dodecyl sulfate). Based on the results of yield, integrity, and purity, we found that the modified CTAB method is more suitable to extract high-quality DNA for deep-sea holothurians, with a large amount of yield, minimized degradation, and higher purity. This study provides a high-quality DNA of deep-sea holothurians for genome sequencing and further studies.KEYWORDS: DNA extractiondeep-sea holothurianshigh-molecular-weight DNACTAB AcknowledgmentsWe thank the captains, crews, and scientific staff on R/V Tansuo 1, and the pilots of HOV Shenhaiyongshi. Special thanks to Dr. El-Hadji Malick Cisse (Hainan University) for improving the English writing in this manuscript.Disclosure statementNo potential conflict of interest was reported by the author(s).Supplementary materialSupplemental data for this article can be accessed online at https://doi.org/10.1080/10236244.2023.2241977.Additional informationFundingThis study was supported by the major scientific and technological projects of Hainan Province [ZDKJ2021036, ZDKJ2019011], National Natural Science Foundation of China [42106121], Strategic Priority Research Program of the Chinese Academy of Sciences [XDA22050303].