Comparative Study on Three Methods for the Extraction of Total RNA from Grape Leaves(PDF)

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:

2013年02期

Page:

87-90

Research Field:

Publishing date:

Info

Title:

Comparative Study on Three Methods for the Extraction of Total RNA from Grape Leaves

Author(s):

YANG Xiao-yan¹; ZHANG Bo¹; 2; HUANG Fang-ai¹; YAN Huan¹; LI Yue-rong¹

1.School of Pharmaceutical Sciences，Shihezi University，Shihezi，Xinjiang 832002；

2.Constructing Key Lab by Province and MOST of Xinjiang Special Plant Resources，Shihezi，Xinjiang 832000

Keywords:

grape leaves of Vitis vinifera L.cv.Red Globe; total RNA; LiCl precipitation method; extraction methods

PACS:

S 663.1

DOI:

-

Abstract:

Three extraction methods of RNeasy Mini Kit，hot phenol method and LiCl precipitation were used to extract total RNA from grape leaves，and their integrity and purity were detected by via agarose gel electrophoresis and Agilent 2100 analyser for a comparative study，in order to compare three RNA extraction methods and find out the suitable one for isolating intact and high quality total RNA from grape leaves (Vitis vinifera L.cv.Red Globe).The results showed that among the three extraction methods，LiCl precipitation method demonstrated higher yield and better integrity of total extracted RNA，with a OD₂₆₀/OD₂₈₀ ratio between 1.8~2.0 and clear 28S and 18S bands in electrophoresis pattern.It could meet the need in molecular biological studies.

References:

[1]王文锋,肖月华,侯磊,等.棉花总RNA的快速提取方法[J].河南农业大学报,2002,36(3):229-231.

[2]胡国斌,梅兴国,刘怡.改良异硫氰酸胍一步法提取红豆杉细胞RNA[J].生物技术,2001,11(5):31-33.

[3]成建红,张玉刚,李天忠.室温离心SDS法快速提取苹果等植物组织RNA[J].园艺学报,2006,33(3):470.

[4]刘洋,何心尧,马红波,等.用CTAB-PVP法提取棉花各组织总RNA的研究[J].中国农业大学学报,2006,11(1):53-56.

[5]Wan C H，Wilkins T A.A modified hot borate method significantiy enhances the yield of high-quality RNA from cotton(Gossypium hirsutum L.)[J].Anal Biochem,1994,223(1):7-12.

[6]刘长军,孟玉玲,侯嵩生,等.棉花法呢基焦磷酸合酶cDNA克隆、序列分析及其在种子发育过程中的表达特征[J].植物学报,1998,40(8):703-710.

[7]Reid K E，Olsson N，Schlosser J，et al.An optimized grapevine RNA isolation procedure and statistical determination of reference genes for real-time RT-PCR during berry development[J].BMC Plant Biology，2006,6(27):1-11.

[8]赵巍巍,宗成文,曹后男,等.葡萄花序总RNA提取方法研究[J].安徽农业科学,2009,37(32):16161-16162.

[9]张今今,王跃进,王西平,等.葡萄总RNA提取方法的研究[J].果树学报,2003,20(3):178-181.

[10]房经贵,高志红,陶建敏.一种提取葡萄芽中总RNA的方法[J].生物技术,2003(2):24-25.

[11]鲁晓燕,赵英,孙文英.一种有效的葡萄叶片总RNA提取方法[J].新疆农业科学,2004,41(6):456-457.

[12]李西萍,牛建新,张强，等.葡萄茎痘病毒病的RT-PCR检测[J].果树学报,2005,22(6):715-716.

[13]宋红苗,马杰,徐祥彬.一种从富含多糖果实中提取RNA的方法[J].浙江农业科学,2012(1):87-89.

[14]Lewinsohn E，Steel L C，Croteau R.Simple isolation of functional RNA from woody stems of gymnospenns[J].Plant Mol Biol Rep,1994(12):20.

[15]周向红,易乐飞,王萍.桑椹总RNA 抽提方法的比较[J].食品科学,2011,32(12):209-212.

[16]司爱君,阮孟斌,谢宗铭.改良热酚法快速提取棉花不同组织RNA[J].安徽农业科学,2012,40(16):8830-8832.

[17]Wang Y，Li Z，Liu X X，et al．A Comparative Study on Three Methods for the Extraction of Total RNA from Pinus bungeana[J].Agricultural Biotechnology,2011,12(5):663-665.

[18]王暑辉,徐倩,徐筱,等.富含多糖多酚的侧柏叶片总RNA提取方法[J].吉林农业大学学报,2012,34(1):76-80.

[19]赵晓,马会勤,陈尚武,等.葡萄果实发育后期半定量RT-PCR内参基因的优选[J].中国农业大学学报,2010(15):7-14.

[20]Wang C S，Vodkin L O.Extraction of RNA from tissues containing high levels of proeyanidins that bind RNA[J].Plant Molecular Biology Reporter,1994,12(2):132-145.

[21]闫光照,郑根昌,等.β-巯基乙醇与PVP 在红干椒DNA提取中的比较研究[J].内蒙古民族大学学报(自然科学版),2009,24(1):51-53.

[22]Fang G，Hammar S，Grumet R.A quick and inexpensive method for removing polysaccharides from plant genomic DNA[J].Bio Techniques,1992,13(1):52-56.

Memo

Memo:

-

Common functions