Bijan Majumdar

A low-cost disruption of rhizospheric microorganisms for the extraction of total RNA using modified RNeasy Mini Kit protocols

A low-cost disruption of jute (Corchorus olitorius) rhizospheric microorganisms (Aspergillus niger and Arthrobacter sp.) with naturally occurring sand for extracting total RNA using modified QIAGEN RNeasy Mini Kit protocols is described. The method yielded distinct 18S and 28S, and 16S and 23S subunits of the fungal and bacterial ribosomal RNAs, respectively, on formaldehyde agarose gels. Total RNA extracted using this method was suitable for reverse transcription PCR to study the differential expression of mRNAs under the stress of herbicides. Primers OPM 1 and OPP 15 were effective in identifying upregulated transcripts in glyphosate and paraquat stressed A. niger. Similarly, primers OPP 13 and OPR 7 were effective in identifying upregulated transcripts in glyphosate-stressed Arthrobacter sp. This is the first report to show that natural sand can be combined with QIAGEN RNeasy Mini Kits for the extraction of total RNA from rhizospheric microorganisms. Novelty of the present method lies in the grinding of samples without the need of liquid nitrogen and then direct purification of RNA using modified QIAGEN RNeasy Mini Kit protocols, without chloroform extraction.