|Table of Contents|

Extraction of High-quality Mitochondrial DNA of Prunus persica for Genome Sequencing

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2019年18
Page:
19-25
Research Field:
Publishing date:

Info

Title:
Extraction of High-quality Mitochondrial DNA of Prunus persica for Genome Sequencing
Author(s):
ZHANG Ying; WU Junkai; WANG Haijing; LIU Luqi; YIN Yarui; ZHANG Libin
(College of Horticulture Science ＆ Technology，Hebei Normal University of Science ＆ Technology，Qinhuangdao,Hebei 066000)
Keywords:
Prunus persica; mitochondrial DNA extraction; callus; high throughput sequencing
PACS:
-
DOI:
10.11937/bfyy.20190333
Abstract:
High quality of mitochondrial DNA (mtDNA) is a fundamental basis for mitochondrial genome sequencing and sequencing analysis.Callus from ‘21st Century’peach leaves were induced at 24 ℃ under completely dark conditions callus induction and proliferution.The obtained callus was homogenized and mtDNA was extracted and purified by differential centrifugation，sucrose liner and SDS pyrolysis.The results showed that,the callus，of which the induction rate could reach 100%，could be effectively induced and proliferated with 3.0 mg?L-1 6-BA and 1.0 mg?L-1 2,4-D.The purified mitochondria,detected by fluorescence microscopy,were complete and active.The yield of purified mtDNA extracted from callus was 13.10 μg?g-1 without RNA and proteins.The band size of mtDNA electrophoresis was about 23 kb without degradation.The obtained mtDNA was free of nuclear DNA and chloroplast DNA contamination by specific gene PCR.In this study，a rapid and efficient extraction technology system suitable for high quality of mitochondrial DNA in Prunus persica was established.It laid a reference for the subsequent research on relevant molecular biology.

References:

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Last Update: 2019-09-29