RNA. Study and Optimization of a Method for High2Quality RNA Extraction From Pa nax ginseng Root Tissues. E2mail : sina.

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1 2010,32 (5) : ,522 http :/ / xuebao. jlau. edu. cn Journal of Jilin Agricultural University E2mail : sina. com RNA Ξ 1, 2, 1, 2ΞΞ 1., ; 2., : CTAB2LiCl pbiozol RNAout Trizol 4 RNA :4 RNA Trizol RNA, RNA, RNA g,od 260 / OD , OD 260 / OD : RNA ; ; : R28412 : A : (2010) Study and Optimization of a Method for High2Quality RNA Extraction From Pa nax ginseng Root Tissues HU Wei 1, BAI Shi 2, ZHU Tao 1, ZHANG Lian2xue 2 1. College of Life Science, Jilin Agricultural University, Changchun , China ; 2. College of Chi2 nese Medicinal Materials, Jilin Agricultural University, Changchun , China Abstract : Extraction of high2quality RNA from Panax ginseng root tissues is very difficult due to high levels of polyphenolics and polysaccharides that bind or coprecipitate with RNA. The methods such as CTAB2LiCl, pbiozol, RNAout and optimized Trizol were applied to isolate high purity and integrity RNA from Panax ginseng root. By all of these methods total RNA can be obtained, but only by the opti2 mized Trizol method, it extracts integrity and high class RNA from mature root tissue. RNA production rate for g/ g total RNA from root tissues can be obtained. The absorbance ratios (OD 260 / OD 280 ) were 118 to 210. The absorbance ratios (OD 260 / OD 230 ) were 210 to 213. Key words : RNA extraction ; Panax ginseng ; condition optimization ( Panax ginseng), [1 ],,(gin2 senoside), [223 ],, [425 ], RNA Ξ ΞΞ : ( ) :,,,, : : :

2 5 06 RNA,,,, RNA, [6 ], RNA, RNA 4 RNA, RNA Trizol, RNA, 1 111, 70 %1 min,,, 011 % DEPC h, h 3 % H 2 O Trizol Invitrogen,RNAout,pBIOZOL RNA,PVP DEPC AMRESCO, CTAB2LiCl RNA 100 mg,,015 D, 115 ml 1 ml CTAB,65 30 min 115 ml,,65 10 min,4 112 r/ min 10 min,,, 10 min,112 r/ min 10 min, 2 3,1/ 3 LiCl (8 mol/ L),4 4, 112 r/ min 20 min, 70 % 2,DEPC (20 L) RNA, pbiozol RNA 100 mg, 015 ml (4 ) pbio2 ZOL,, 5 min 112 r/ min 2 min,011 ml NaCl (5 mol/ L), 013 ml,, r/ min 10 min,, 10 min, r/ min 10 min, 1 ml 75 %, 112 r/ min 1 min,30 L DEPC RNA RNAout RNA A mg 1 ml A, 115 ml, 30 s013 ml B 012 ml, 30 s, 113 r/ min 5 min( 800 L) 115 ml, DNA,, 100 L 500 L C 200 L, 30 s, 113 r/ min 3 min, 115 ml 30 s,, 113 r/ min 3 5 min,rna 75 %1 ml, 30 s, 113 r/ min 1 min,, RNA 20 L DEPC RNA, Trizol RNA mg,,, 1 ml Trizol 115 ml,, 10 min ;4 112 r/ min 10 min,

3 ,;200 L 50 L (2 mol/ L, ph412) 15 L 2 1 %PVP 10 L,,10 min ;4 112 r/ min 15 min, ;( 015 ml) 200 L (3 mol/ L),, h ;4,DNA, 112 r/ min 10 min, ; 1 ml 75 % ;4 112 r/ min 5 min,, 10 min,20 L DEPC RNA, RNA 110 %RNA TU ( ) RNA 230,260, 280 nm, 1 L RNA,1 ml DEPC, RNA OD 260 / OD OD 260 / OD RNA (OD 260 RNA 40 g/ ml) RNA : RNA ( g/ g) = [ OD g/ ml V RNA (ml) ]/ RNA 4 RNA 1 % 1 1,4 RNA, CTAB2LiCl RNA,28S 18S,, RNA ;RNAout RNA 3 : RNA 507, 28S 18S 2 1 ;pbio2 ZOL RNA 3,, ; Trizol RNA 28S 18S 2,2 1, RNA, a. CTAB2LiCl The CTAB2LiCl method ;b. RNAout The RNAout method ;c. pbiozol The pbiozol method ; d. Trizol The optimized trizol method 1 4 RNA Fig. 1. The electrophore sis analysis of total RNA isola2 tion from Panax ginseng root tissue s with dif2 ferent methods 212 RNA RNA RNA OD 260 / 0D , < 118, ; > 210, 2 RNA OD 260 / OD , 1 :Trizol RNA 1 4 RNA Table 1. The detection of the quality of total RNA from Panax ginseng root by four methods Method OD 260 nm 280 nm 230 nm OD 260 / OD 280 OD 260 / OD / ( g g - 1 ) 230 Yield CTAB2LiCl CTAB2LiCl method RNAout RNAoutl method pbiozol pbiozol method Trizol Optimized trizol method Journal of Jilin Agricultural University

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