A simple and inexpensive protocol for DNA isolation from avian blood
PDF
PDF

Keywords

DNA Isolation
Avian Blood
Simple
Inexpensive
Vultures.

How to Cite

Kulkarni, M. D., Shringarpure, R., Bhatt, P., Prakash, N., & Prakash, V. (2015). A simple and inexpensive protocol for DNA isolation from avian blood. Journal of the Bombay Natural History Society (JBNHS), 112(1), 3–7. https://doi.org/10.17087/jbnhs/2015/v112i1/92186

Abstract

One of the many challenges encountered during species conservation programmes is genetic management of threatened populations. Declining populations face the threat of genetic drift and/or imbalance in sex ratios, adding to the risk of extinction. The primary step in genetic management of any population is the isolation of DNA from an available tissue sample. Good quality and quantity of DNA, whenever isolated, could be preserved and applied in a number of studies. Collection of blood, which is a very good source of DNA, from birds is possible in conservation programmes. A simple and inexpensive protocol for DNA isolation facilitates processing the blood samples in the field with ease. Commercial kits, though available, are expensive and have limited shelf life. Herein, we report a simple protocol of DNA isolation from avian blood. The protocol did not employ any proteases or organic solvent, making it comparatively inexpensive. This protocol, initially developed for vulture blood, was successfully applied later for other species. The protocol could isolate DNA sufficient for at least 50 amplification reactions. The isolated DNA was found suitable for spectrometry as well as downstream applications like PCR and cloning.
https://doi.org/10.17087/jbnhs/2015/v112i1/92186

References

Arshad M., J. Gonzalez, A.A. El-Sayed, T. Osb orne & M. Wink (2009): Phylogeny and phylogeography of critically endangered Gyps species based on nuclear and mitochondrial markers. Journal of Ornithology 150(2): 419-430.

Bailes, S.M., J.J. Devers, J.D. Kirby & D.D. Rhoads (2007): An inexpensive, simple protocol for DNA isolation from blood for high-throughput genotyping by polymerase chain reaction or restriction endonuclease digestion. Poultry Science 86(1): 102-106.

Banhos, A., T. Hrbek, W. Gravena, T. Sanaiotti & I.P. Farias (2008): Genomic resources for the conservation and management of the Harpy Eagle (Harpia harpyja, Falconiformes, Accipitridae). Genetics and Molecular Biology 31: 146-154.

Belovsky, G.E., C. Mellison, C. Larson & P.A. Van Zandt (1999): Experimental studies of extinction dynamics. Science 286 (5442): 1175-1177.

Breininger, D.R., M.A. Brugman & B.M. Stith (1999): Influence of habitat quality, catastrophes, and population size on extinction risk of the Florida scrub-jay. Wildlife Society Bulletin 27: 810-822.

Chou, T.C., C.T. Yao, S.H. Su, Y.C. Hung, W.S. Chen, C.C. Cheng, C.N. Tseng, H.M. Wang, Y.C. Chou, S.S. Li, D.L. Gu & H.W. Chang (2010): Validation of Spilornis cheela hoya TaqMan probes for potential gender identification of many Accipitridae species. Theriogenology 73(3): 404-411.

Fischer, M. & J. Stock lin (1997): Local extinctions of plants in remnants of extensively used calcareous grasslands 1950-1985. Conservation Biology 11(3): 727-737.

Geyer, C.J., O.A. Ryder, L.G. Chemnick & E.A. Thompson (1993): Analysis of relatedness in the California condors, from DNA fingerprints. Molecular Biology & Evolution 10(3): 571-589.

Ghorpade, P.B., P.K. Gupta, V. Prakash, R.J. Cuthbert, M. Kulkarni, N. Prakash, A. Das, A.K. Sharma & M. Saini (2012): Molecular sexing of threatened Gyps vultures: an important strategy for conservation breeding and ecological studies. SpringerPlus 1: 62.

Griffiths, R., M.C. Double, K. Orr & R.G.J. Daws on (1998): A DNA test to sex most birds. Molecular Ecology 7: 1071-1075.

Grimb erg, J., S. Nawosc hik, L. Ellusc io, R. McKee, A. Turck ,& A. Eisenberg (1989): A simple and efficient non-organic procedure for the isolation of genomic DNA from blood. Nucleic acid Research 17: 83-90.

Haig, S.M., J.D. Ballou & S.R. Derricks on (1990): Management options for preserving genetic diversity: reintroduction of Guam Rails to the wild. Conservation Biology 4(3): 290-300.

Helms , C. (2002): Salting out Procedure for Human DNA extraction. In: The Donis-Keller Lab - Lab Manual Homepage. Available from:http://www.protocol-online.org/prot/Protocols/Extractionofgenomic-DNA-from-whole-blood-3171.html.

IUCN (2015): IUCN Red List of Threatened Species. Version 2015.4 (www.iucnredlist.org).

Khosravinia H., H.N. Narasimha Murthy, D. Thertha Parasad & N. Pirany (2007): Optimizing factors influencing DNA extraction from fresh whole avian blood. African Journal of Biotechnology 6(4): 481-486.

Kulkarni, M.D., R. Shringarpure, S. Chandramohan, M. Saini, T.K. Palai, P.K. Gupta, P. Bhatt, N. Prakash, A. Bhagwat & V. Prakash (2014): Molecular sexing of the critically endangered Slender-billed Vulture Gyps tenuirostris and its conservation implications. J. Bombay Nat. Hist. Soc. 111(2): 73-80.

Malago, W.J., H. Franco, E.J. Matheucc i, A. Medaglia & F. Henrique Silva (2002): Large scale sex typing of ostriches using DNA extracted from feathers. BMC Biotechnology 2(1): 19.

Miller, B., D. Biggins, L. Hanebury & A. Vargas (1994): Reintroduction of the black-footed ferret (Mustela nigripes). Pp. 455-464. In: Olney P.J.S., G.M. Mace & A.T.C. Feistner (Eds): Creative Conservation - Interactive management of wild and captive animals. Chapman & Hall, London.

Newmark, W.D. (1995): Extinction of mammal populations in western North-American National Parks. Conservation Biology 9(3): 512-526.

Rudbeck , L. & J. Diss ing (1998): Rapid, simple alkaline extraction of human genomic DNA from whole blood, buccal epithelial cells, semen and forensic stains for PCR. Biotechniques 25(4): 588-592.

Sambrook, J., E.F. Fritsch & T. Maniatis (1989): Molecular Cloning: A Laboratory Manual. Vol. 1. 2nd edn. Cold Spring Harbor Laboratory Press.

Yokota, M., K. Sindo, M. Hiyoshi, I. Tsuda & N. Tatsumi (1998): A convenient method of DNA extraction from blood anticoagulated with EDTA. Biochemistry and Molecular Biology International 45(3): 617-622.

Downloads

Download data is not yet available.