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Optimization of DNA isolation from legume nodules
Author(s) -
KrasovaWade T.,
Neyra M.
Publication year - 2007
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2007.02149.x
Subject(s) - legume , vigna , dna , thiocyanate , guanidine , dna extraction , root nodule , biology , nodule (geology) , yield (engineering) , genomic dna , fabaceae , botany , chromatography , microbiology and biotechnology , chemistry , biochemistry , polymerase chain reaction , bacteria , genetics , symbiosis , gene , materials science , paleontology , metallurgy
Aims: The aim of this study was to optimize DNA extraction from legume nodules to obtain large amounts of high‐quality genomic DNA. Methods and Results: Nodules of different legume species were used. Varied concentrations of guanidine thiocyanate (from 6 mol l −1 to 0·05 mmol l −1 ), a component of DNAzol, were tested. The quality of DNA extract was determined by PCR–RFLP. The best results were obtained with 0·5 mmol l −1 guanidine thiocyanate, which resulted in greater DNA yield than with higher and lower concentrations or with DNAzol. Conclusion: The procedure using 0·5 mmol l −1 guanidine thiocyanate yields the highest DNA amount when compared with previously described protocols and offers a reliable method to isolate DNA from nodules of different origins. Significance and Impact of the Study: Irrespective of nodule origin, DNA yield was increased significantly, by two (e.g., Vigna nodules) to seven ( Acacia auricoliformis nodules) times. In addition, the proposed procedure’s costs are lower than those using the DNAzol.