Comparison of Total RNA Extraction Methods from Nervilia fordii (Hance) Schltr.Leaves(PDF)

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:

2013年02期

Page:

91-94

Research Field:

Publishing date:

Info

Title:

Comparison of Total RNA Extraction Methods from Nervilia fordii (Hance) Schltr.Leaves

Author(s):

LIANG Ling-ling¹; DENG Yu-xing²; HUANG Qiong-lin¹; HE Rui¹; ZHAN Ruo-ting¹

1.Key Laboratory of Chinese Medicinal Resources from Lingnan，Ministry of Education，Research Center of Chinese Medicinal Resource Science and Engineering，Guangzhou University of Traditional Chinese Medicine，Guangzhou，Guangdong 510006；

2.School of Chinese Materia Medica，Guangzhou University of Traditional Chinese Medicine，Guangzhou，Guangdong 510006

Keywords:

Nervilia fordii (Hance) Schltr.; total RNA extraction; RT-PCR

PACS:

S 567.23+9

DOI:

-

Abstract:

Five different extraction methods(RNeasy Blant Miri kit,RNAiso Plus,RNAiso for Polysaccharide-rich Plant Tissue,CTAB and SDS)，combined with four different extraction solutions were used to extract total RNA from Nervilia fordii (Hance) Schltr.leaves.The effect and efficiency of extraction was evaluated by agarose gel electrophoresis，ultraviolet spectrophotometry and RT-PCR.The results showed that apart from RNAiso Plus method，total RNA resulted from the other methods all showed genomic DNA pollution.Among these methods，the total RNA resulted from RNAiso Plus method followed with extraction solution ③(1/5KAC+4/5PCI) was the most integrated，and the total RNA resulted from SDS method followed with extraction solution ④(1/5NaCl+4/5PCI) had the highest concentration (372.4 ng/μL).Total RNA obtained from twenty treatment groups have been used to successfully amplify 18s rRNA gene segment by RT-PCR.This study established a proper total RNA extraction method for Nervilia fordii (Hance) Schltr.leaves，and it provided technique basis for gene cloning，expression，and transcriptomic analysis of the plant.

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