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Simultaneous isolation of DNA, RNA, and protein from Medicago truncatula L.
Author(s) -
Xiong Junbo,
Yang Qingchuan,
Kang Junmei,
Sun Yan,
Zhang Tiejun,
Margaret Gruber,
Ding Wang
Publication year - 2011
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201000425
Subject(s) - trizol , nucleic acid , medicago truncatula , chromatography , trichloroacetic acid , sample preparation , chemistry , rna , phenol extraction , extraction (chemistry) , dna extraction , rna extraction , protein purification , biochemistry , biology , gene , polymerase chain reaction , bacteria , genetics , symbiosis
We describe a method for the simultaneous extraction of proteins and nucleic acids from Medicago truncatula tissues. Using a modified TRIzol reagent method, we developed a simple and an effective way to simultaneously extract proteins and nucleic acids from a single sample. We verified that this method does not affect the quality or quantitation of the isolated DNA and RNA. Furthermore, we used 2‐DE to compare M. truncatula leaf, stem, and root samples processed using this new method with two commonly used methods: phenol extraction/methanol–ammonium acetate precipitation and trichloroacetic acid/acetone precipitation. The results showed that our method was superior to the other methods, based on 2‐DE patterns. We also demonstrated that our protocol is compatible with proteomic analysis, as 10 out of 14 selected proteins isolated by the method were identified by MALDI‐TOF‐MS/MS. The protocol described can be used with sample preparation protocols for proteomic, transcriptomic, and genomic studies.