Extraction of Genomic DNA from Dry Leaves of by Modified CTAB Methods

WANG Gui-e; CHAO Qun-fang; LIANG Jian-fang; YANG Yang; YIN Ya-lan

doi:10.13422/j.cnki.syfjx.2015120019

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Extraction of Genomic DNA from Dry Leaves of by Modified CTAB Methods

更新时间：2024-01-04

- Extraction of Genomic DNA from Dry Leaves of by Modified CTAB Methods
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 21, Issue 12, Pages: 19-22(2015)
- 作者机构：
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- DOI：10.13422/j.cnki.syfjx.2015120019
  CLC： R284.2
- Published：2015
- 稿件说明：
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WANG Gui-e, CHAO Qun-fang, LIANG Jian-fang, et al. Extraction of Genomic DNA from Dry Leaves of by Modified CTAB Methods[J]. Chinese journal of experimental traditional medical formulae, 2015, 21(12): 19-22.
DOI：

WANG Gui-e, CHAO Qun-fang, LIANG Jian-fang, et al. Extraction of Genomic DNA from Dry Leaves of by Modified CTAB Methods[J]. Chinese journal of experimental traditional medical formulae, 2015, 21(12): 19-22. DOI： 10.13422/j.cnki.syfjx.2015120019.

摘要

目的: 从自然干燥的新疆一枝蒿中提取高质量的基因组DNA

为该药材的分子生物学研究提供参考。方法: 采用2种改良十六烷基三甲基溴化铵(CTAB)法与常规CTAB法提取新疆一枝蒿中基因组DNA

通过加适量聚乙烯吡咯烷酮(PVP)研磨样本

在核裂解前用细胞核裂解液(STE)洗涤多酚类和多糖类物质

利用95%乙醇室温沉淀DNA等措施。改良CTAB法2与1相比不用液氮研磨样本。通过琼脂糖凝胶电泳、紫外分光光度法和相关序列扩增多态性(SRAP)检测提取DNA的质量。结果: 2种改良CTAB法所得DNA质量均优于常规CTAB法

DNA完整性较好

A260 nm/A280 nm在1.8~2.0

多糖类杂质去除较为干净。SRAP检验条带清晰、多态性良好。结论: 2种改良CTAB法均适用于高质量新疆一枝蒿干叶基因组DNA的提取

得到的DNA纯度和完整性均较理想

可为后续分子生物学研究提供良好模版。

Abstract

Objective: To extract high quality genomic DNA from dry leaves of Artemisia rupestris dried naturally and lie a foundation for further molecular experiments. Method: Three different methods

conventional hexadecyl trimethyl ammonium bromide (CTAB) method and modified CTAB methods were applied to isolate genomic DNA from dry leaves of A. rupestris.Two modified CTAB methods were different from conventional CTAB method in three ways

nucleus lysate(STE) was used for removal of impurities

polyvinyl pyrrolidone (PVP) was added when grinding samples and 95% ethanol for precipition of DNA in room temperature.These two modified CTAB methods were different from whether liquid nitrogen was added while grinding samples.Purity and integrality of DNA were detected by UV spectrophotometry

agarose gel electrophoresis and sequence-related amplified polymorphism (SRAP). Result: Higher purity DNA was obtained by improved CTAB methods by comapred with conventional CTAB method

value of A260 nm/A280 nmwas between 1.8-2.0

polysaccharides was removed relatively clear.SRAP produced clear polymorphic patterns. Conclusion: These two modified CTAB methods are suitable for DNA extraction from dry leaves of A. rupestris

purity and integrality of isolated DNA can be used in further molecular biology research.

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