Multiphase modification of Tissue Lyser Technique based Genomic DNA Isolation Protocol in Finger Millet (Eleusine coracana L)

Author: Prafull Kumar, A K Sarawgi, B.K. Das, Deepak Sharma, Gautam Vishwakarma and Vikash Kumar

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Abstract

The semi-cereal crop Finger millet (Eleusine coracana (L) Gaertn) is presently gaining identity for its nutraceutical and climate resilient sustainable nature but research acceleration is being limited by non-application of high-throughput molecular biology protocols. 210 finger millet genotypes were taken by deploying time saving tissue lyser based DNA isolation technique for further molecular studies. Due to new crop biology viz., C4 carbon fixation pathway and presence of complex biochemical compounds like phenols; and non-parallelism with other model plant species’ a new crop specific DNA isolation protocol termed “easy step” was optimized. Utilizing standard reagents and chemicals of CTAB method of DNA isolation for plants, different aspect of DNA isolation viz. contamination with phenols, proper disintegration and high efficiency purification were optimized. Different combination, tissue lyser oscillation frequency and duration, along with other purification steps were opti

Keywords

Phenolic compounds, PVP, Tissue lyser, C4 crop, Finger millet.

Conclusion

Involvement of substantial amount of time and laboratory resources persuaded us to summarize and recompile the work as manuscript. Although, initially the report can be taken casually due to well standardised in other crop species namely rice, wheat, maize, oilseeds, pulses etc.; but in Finger millet the sophisticated molecular biology tools are yet to be explored with fine tune. We suggest from the study that taking 10-15 days old seedling using comb-stand method, addition of 1 percent (v/v) of Polyvinylpyrrolidone (PVP) complemented CTAB extraction buffer, using tissue lyser for crushing of leaf samples for 150 seconds at the interval of every 30 seconds and above discussed combination of temperature, spinning and timing for isolation of quality DNA in this limited explored crop species. The high quality and rapid DNA isolate will enrich the way of E. coracana towards high throughput techniques like next generation sequencing (Nguyen et al., 2019). Through combining the efforts of mo

References

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How to cite this article

Kumar, P., Sarawgi, A.K., Das, B.K., Sharma, D., Vishwakarma, G. and Kumar, V. (2021). Multiphase modification of Tissue Lyser Technique based Genomic DNA isolation protocol in finger millet (Eleusine coracana L). Biological Forum – An International