TY - JOUR
T1 - An efficient method for extraction of RNA from rice leaves at different ages using benzyl chloride
AU - Suzuki, Y.
AU - Makino, A.
AU - Mae, T.
N1 - Funding Information:
We thank Drs Ko Iba and Kensuke Kusumi (Kyushu University) for a kind gift of cDNA clones of rice and their advice on the experiment. We also thank Drs Tomoyuki Yamaya and Toshihiko Hayakawa and members of their laboratories for their instruction and fruitful discussion and for use of their equipment. This work was supported by Grants-in-Aid for Science Research 12460028 and for Science Research in Priority Areas 12025202 from the Ministry of Education, Science and Culture of Japan, by Grant-JSPS-RFTF 96L00604 for Research for the Future from the Japan Society for the Promotion of Science, and by the Bio Design Program (BDP-01-I-1-1) from the Ministry of Agriculture, Forestry and Fisheries, Japan.
PY - 2001
Y1 - 2001
N2 - With a conventional method of RNA extraction using an acid guanidinium thiocyanate-water-saturated phenol-chloroform mixture, extraction efficiency of extractable RNA to total RNA (extractable RNA+ residual RNA) in rice leaves at various ages was 54-69%. With a new method, an improvement of the above, using benzyl chloride instead of water-saturated phenol together with further maceration with a small amount of quartz sand, the efficiency was increased to 81-95%. When RNA fractions obtained with the improved method were subjected to agarose gel electrophoresis, intact bands of 25 S and 17 S rRNAs were detected. With a DNA probe for rice rbcS, only a single band was observed on the blotted membrane. These results indicate that the improved extraction method of RNA with benzyl chloride is useful for quantitative and qualitative analysis of RNA in plant tissues such as stiff leaves of rice.
AB - With a conventional method of RNA extraction using an acid guanidinium thiocyanate-water-saturated phenol-chloroform mixture, extraction efficiency of extractable RNA to total RNA (extractable RNA+ residual RNA) in rice leaves at various ages was 54-69%. With a new method, an improvement of the above, using benzyl chloride instead of water-saturated phenol together with further maceration with a small amount of quartz sand, the efficiency was increased to 81-95%. When RNA fractions obtained with the improved method were subjected to agarose gel electrophoresis, intact bands of 25 S and 17 S rRNAs were detected. With a DNA probe for rice rbcS, only a single band was observed on the blotted membrane. These results indicate that the improved extraction method of RNA with benzyl chloride is useful for quantitative and qualitative analysis of RNA in plant tissues such as stiff leaves of rice.
KW - Oryza sativa L.
KW - RNA extraction
KW - Senescence
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U2 - 10.1093/jexbot/52.360.1575
DO - 10.1093/jexbot/52.360.1575
M3 - Article
C2 - 11457918
AN - SCOPUS:0034942046
SN - 0022-0957
VL - 52
SP - 1575
EP - 1579
JO - Journal of Experimental Botany
JF - Journal of Experimental Botany
IS - 360
ER -