Novel and PCR ready rapid DNA isolation from Drosophila

Abstract

Introduction

Isolation of genomic DNA is an initial step in molecular biology techniques. The quality of isolated DNA depends on procedures and chemicals, as well as source and types of the sample used. Several existing procedures are expensive and time consuming. In this study, we isolated high quality genomic DNA with an inexpensive and least time consuming procedure using Drosophila melanogaster flies, larvae, and pupae.

Methods

Drosophila melanogaster samples were collected from pre-cultured bottles, and genomic DNA was extracted using a proposed novel and PCR-ready method from three different pools of flies [PF1, PF2, and PF3], similarly from larvae and pupae [PL1, PL2, PL3, PP1, PP2, and PP3, respectively]. Isolated genomic DNA was subjected to PCR amplification with different dilutions using the COI gene and further amplicons were used for RAPD and DNA sequencing.

Results

The high quality of isolated genomic DNA was confirmed by 0.8% agarose gel electrophoresis and the purity and quantity of the DNA isolated from single fly, larva and pupa was similar to the purity and quantity of the DNA isolated using the NucleoSpin^R Tissue kit method. Isolated genomic DNA was successfully amplified when the template was diluted in the ratio of 1:10. Further successful RAPD amplification and sequencing analysis of the COI gene confirms the efficiency of the downstream application of the proposed novel method.

Conclusion

The present Novel and PCR ready rapid DNA isolation method will be potentially beneficial, and it can be successfully used for quick isolation of high molecular weight DNA from Drosophila flies larvae and pupae for DNA barcoding, identification of new species, genotyping, RAPD analysis, etc. Moreover, it can also be easily scaled up for bulk preparations.

KEYWORD:

• Drosophila
• rapid DNA isolation
• PCR
• RAPD
• sequencing

Acknowledgement

We thank Karnataka Institute for DNA Research (KIDNAR), Dharwad for constant support throughout the research. We also thank Ms Pooja Joshi, Ms. Divya Bhat, and Mr. Vishweshwar Bhat for their help during the work.

Disclosure statement

The authors declared no conflicts of interest concerning the research, authorship, funding and publication.

Data availability statement

Not applicable.

Additional information

Funding

This study was financially supported by Karnataka Institute for DNA Research (KIDNAR) Dharwad. Karnataka, India.