干旱区生态环境知识资源中心(Arid): Extraction of high-quality, high-molecular-weight DNA depends heavily on cell homogenization methods in green microalgae

Arid

DOI	10.1002/aps3.11333
	Extraction of high-quality, high-molecular-weight DNA depends heavily on cell homogenization methods in green microalgae
	Stark, Jordan R.; Cardon, Zoe G.; Peredo, Elena L.
通讯作者	Peredo, Elena L.
来源期刊	APPLICATIONS IN PLANT SCIENCES
ISSN	2168-0450
出版年	2020
卷号	8 期号:3
英文摘要	Premise New sequencing technologies have facilitated genomic studies in green microalgae; however, extracting high-quality DNA is often a bottleneck for long-read sequencing. Methods and Results Here, we present a low-cost, highly transferrable method for the extraction of high-molecular-weight (HMW), high-purity DNA from microalgae. We first determined the effect of sample preparation on DNA quality using three homogenization methods: manual grinding using a mini-pestle, automatic grinding using a vortex adapter, and grinding in liquid nitrogen. We demonstrated the versatility of grinding in liquid nitrogen followed by a modified cetyltrimethylammonium bromide (CTAB) extraction across a suite of aquatic- and desert-evolved algal taxa. Finally, we tested the protocol's robustness by doubling the input material to increase yield, producing per sample up to 20 mu g of high-purity DNA longer than 21.2 kbp. Conclusions All homogenization methods produced DNA within acceptable parameters for purity, but only liquid nitrogen grinding resulted in HMW DNA. The optimization of cell lysis while minimizing DNA shearing is therefore crucial for the isolation of DNA for long-read genomic sequencing because template DNA length strongly affects read output and length.
英文关键词	DNA integrity long-read sequencing modified CTAB extraction Scenedesmaceae
类型	Article
语种	英语
国家	USA
开放获取类型	Green Published, gold
收录类别	SCI-E
WOS记录号	WOS:000520981900003
WOS关键词	REMOVAL
WOS类目	Plant Sciences
WOS研究方向	Plant Sciences
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/314036
作者单位	Marine Biol Lab, Ctr Ecosyst, 7 MBL St, Woods Hole, MA 02543 USA
推荐引用方式 GB/T 7714	Stark, Jordan R.,Cardon, Zoe G.,Peredo, Elena L.. Extraction of high-quality, high-molecular-weight DNA depends heavily on cell homogenization methods in green microalgae[J],2020,8(3).
APA	Stark, Jordan R.,Cardon, Zoe G.,&Peredo, Elena L..(2020).Extraction of high-quality, high-molecular-weight DNA depends heavily on cell homogenization methods in green microalgae.APPLICATIONS IN PLANT SCIENCES,8(3).
MLA	Stark, Jordan R.,et al."Extraction of high-quality, high-molecular-weight DNA depends heavily on cell homogenization methods in green microalgae".APPLICATIONS IN PLANT SCIENCES 8.3(2020).