Eggshell membrane for DNA sexing of the endangered Maleo (Macrocephalon maleo) [version 3; peer review: 2 approved]

Background: Noninvasive DNA sampling has been applied across many avian genetic studies for a variety of purposes including conservation and management of endangered birds. However, its application in megapodes is still lacking. The previous genetic studies on megapodes used either blood or fresh ti...

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Main Authors: Pramana Yuda, Andie Wijaya Saputra
Format: Article
Language:English
Published: F1000 Research Ltd 2021-01-01
Series:F1000Research
Online Access:https://f1000research.com/articles/9-599/v3
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author Pramana Yuda
Andie Wijaya Saputra
author_facet Pramana Yuda
Andie Wijaya Saputra
author_sort Pramana Yuda
collection DOAJ
description Background: Noninvasive DNA sampling has been applied across many avian genetic studies for a variety of purposes including conservation and management of endangered birds. However, its application in megapodes is still lacking. The previous genetic studies on megapodes used either blood or fresh tissue. Here we present the first demonstration of the use of eggshell membrane for research on endangered Maleo (Macrocephalon maleo).  Methods: We used 24 post-hatched eggshell membranes collected from two different sites, Tambun and Tanjung Binerean, in North Sulawesi, 12 samples in each. Two different DNA extraction methods: alkaline lysis method and gSYNCTM DNA Extraction Kit were applied.  To determine the sex of Maleo, we utilized PCR-based DNA sexing using CHD genes, with the primer set 2550F/2718R.  Results: We successfully extracted all samples; the mean sample concentration was 267.5 ng/µl (range 47–510.5 ng/µl) and samples were of high purity (A260/280 ratio 1.85±0.03). All samples were used to successfully identified sexes, 9 females and 15 males.  Conclusions: Our research clearly illustrates that eggshell membranes can be used for DNA sexing and open the possibility to build noninvasive DNA collections over large spatial scales for population study of endangered birds.
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spelling doaj.art-88dfd6bc3d584afdbeb49bd308769d572022-12-21T19:55:50ZengF1000 Research LtdF1000Research2046-14022021-01-01910.12688/f1000research.23712.331413Eggshell membrane for DNA sexing of the endangered Maleo (Macrocephalon maleo) [version 3; peer review: 2 approved]Pramana Yuda0Andie Wijaya Saputra1Teknobiologi, Universitas Atma Jaya Yogyakarta, Kab. Sleman, DI Yogyakarta, 55281, IndonesiaTeknobiologi, Universitas Atma Jaya Yogyakarta, Kab. Sleman, DI Yogyakarta, 55281, IndonesiaBackground: Noninvasive DNA sampling has been applied across many avian genetic studies for a variety of purposes including conservation and management of endangered birds. However, its application in megapodes is still lacking. The previous genetic studies on megapodes used either blood or fresh tissue. Here we present the first demonstration of the use of eggshell membrane for research on endangered Maleo (Macrocephalon maleo).  Methods: We used 24 post-hatched eggshell membranes collected from two different sites, Tambun and Tanjung Binerean, in North Sulawesi, 12 samples in each. Two different DNA extraction methods: alkaline lysis method and gSYNCTM DNA Extraction Kit were applied.  To determine the sex of Maleo, we utilized PCR-based DNA sexing using CHD genes, with the primer set 2550F/2718R.  Results: We successfully extracted all samples; the mean sample concentration was 267.5 ng/µl (range 47–510.5 ng/µl) and samples were of high purity (A260/280 ratio 1.85±0.03). All samples were used to successfully identified sexes, 9 females and 15 males.  Conclusions: Our research clearly illustrates that eggshell membranes can be used for DNA sexing and open the possibility to build noninvasive DNA collections over large spatial scales for population study of endangered birds.https://f1000research.com/articles/9-599/v3
spellingShingle Pramana Yuda
Andie Wijaya Saputra
Eggshell membrane for DNA sexing of the endangered Maleo (Macrocephalon maleo) [version 3; peer review: 2 approved]
F1000Research
title Eggshell membrane for DNA sexing of the endangered Maleo (Macrocephalon maleo) [version 3; peer review: 2 approved]
title_full Eggshell membrane for DNA sexing of the endangered Maleo (Macrocephalon maleo) [version 3; peer review: 2 approved]
title_fullStr Eggshell membrane for DNA sexing of the endangered Maleo (Macrocephalon maleo) [version 3; peer review: 2 approved]
title_full_unstemmed Eggshell membrane for DNA sexing of the endangered Maleo (Macrocephalon maleo) [version 3; peer review: 2 approved]
title_short Eggshell membrane for DNA sexing of the endangered Maleo (Macrocephalon maleo) [version 3; peer review: 2 approved]
title_sort eggshell membrane for dna sexing of the endangered maleo macrocephalon maleo version 3 peer review 2 approved
url https://f1000research.com/articles/9-599/v3
work_keys_str_mv AT pramanayuda eggshellmembranefordnasexingoftheendangeredmaleomacrocephalonmaleoversion3peerreview2approved
AT andiewijayasaputra eggshellmembranefordnasexingoftheendangeredmaleomacrocephalonmaleoversion3peerreview2approved