Simultaneous isolation of DNA, RNA, and protein from Medicago truncatula L.
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.
Institute of Animal Science, Chinese Academy of Agricultural Science, Yuan Ming Yuan, Beijing, P. R. China.
Junbo
J
Xiong
Qingchuan
Q
Yang
Junmei
J
Kang
Yan
Y
Sun
Tiejun
T
Zhang
Gruber
G
Margaret
Wang
W
Ding
D018744:DNA, Plant / Q000032:analysis / Q000302:isolation & purification*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D046913:Medicago truncatula / Q000737:chemistry*; D018515:Plant Leaves / Q000737:chemistry; D010940:Plant Proteins / Q000032:analysis / Q000302:isolation & purification*; D018517:Plant Roots / Q000737:chemistry; D018547:Plant Stems / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D018749:RNA, Plant / Q000032:analysis / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D018744:DNA, Plant; D010940:Plant Proteins; D018749:RNA, Plant