Standardisation of High Quality RNA Extraction Method for Allium species

Kuldip Jayaswall, Abhishek Bhandawat, Vijay Mahajan, Ashwini Benke, Ram Yashwant Bhomble, Major Singh

Abstract


Secondary metabolites interfere and obstruct the extraction of RNA in many plants including Allium species. Standard protocol for isolation of quality RNA from various tissues of onion and garlic is lacking. A fast and reliable system for isolation of RNA from onion bulbs and garlic cloves is necessary to smooth the progress in functional genomics, transcriptomics, and gene expression studies in garlic & onion. In the current study, we compared two most popular RNA isolation protocols in Allium species for standardization of RNA extraction methods to obtain high quality RNA from Allium species. Among these, the guanidinium-free IHBT-protocol was found to be more suitable for isolation of good quality RNA from Allium tissues, compared to guanidinium-based TRIzol method(TRIzol® (Invitrogen, USA). The isolated RNA was successfully utilized for cDNA synthesis, thus confirming high quality of extracted RNA.

Keywords


Allium, Garlic, Guanidinium-based, Onion, RNA isolation, Secondary metabolites

Full Text:

PDF

References


Chomczynski, P, &Sacchi, N.(1987). Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem,162:156-159.

Bugos, R.C., Chiang, V.L., Zhang, X.H., Campbell, E.R., Podila, G.K.,& Campbell, W.H.(1995).RNA isolation from plant tissues recalcitrant to extraction in guanidine. Bio Techniques,19:734-737.

Ghawana, S., Singh. K., Raizada, J., Rani, A., Bhardwaj. P., Kumar, S.(2007).A method for rapid isolation of RNA and a kit thereof. International Publication No. WO 2007/113614 2007.

Ghawana, S.,Paul, A., Kumar, H.,Kumar, A., Singh, H., Bhardwaj, P.K., Rani, A., Singh, R.S., Raizada, J.,Singh, K.,&Kumar, S.(2011). AnRNA isolation system for plant tissues rich in secondary metabolites. BMC Research Notes.4:85.

Bhardwaj, P.K., Ahuja, P.S., Kumar, S. (2010) Characterization of gene expression of QM from Caraganajubata, a plant species that grows under extreme cold. MolBiol Rep.37:1003-1010.

Dawson, R.M.C., Elliott, D.C., Elliott, W.H., Jones, & K.M. (1986) Data for Biochemical Research. Oxford Science Publication.

Wallace, D.M.(1987) Large and small-scale phenol extractions. Methods Enzymol, 152 (1987): 33-41.


Refbacks

  • There are currently no refbacks.


Online ISSN: 2581-9127

 

Couldn't lock the file.