High-Performance PCR for Alleles Discrimination of Chromo-Helicase-DNA Binding Protein (CHD1) Gene in Bird Sexing
Genetic analyses aiming at assessing the presence of specific sequences or alleles are often carried out by PCR. Sexing of most birds is nowadays based on PCR with “universal” primers and relies on the assessment of the presence of the sex-linked CHD1-Z and -W alleles. The entire workflow is relativ...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-02-01
|
| Series: | Biology |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2079-7737/12/2/300 |
| _version_ | 1797622271040290816 |
|---|---|
| author | Marcello Tagliavia Valentina Catania Giacomo Dell’Omo Bruno Massa |
| author_facet | Marcello Tagliavia Valentina Catania Giacomo Dell’Omo Bruno Massa |
| author_sort | Marcello Tagliavia |
| collection | DOAJ |
| description | Genetic analyses aiming at assessing the presence of specific sequences or alleles are often carried out by PCR. Sexing of most birds is nowadays based on PCR with “universal” primers and relies on the assessment of the presence of the sex-linked CHD1-Z and -W alleles. The entire workflow is relatively time-consuming, especially for batch analyses, whereas methods that allow carrying out the entire procedure in a short time are highly desirable. The only method for outdoor analyses reported so far relies on LAMP; however; it fails to work properly in Procellariiformes. Besides improving the LAMP test; we have developed a PCR-based DNA amplification procedure (named high-performance PCR); whose unique features allow it to outperform standard PCR; making possible the direct, in-tube visual reading of results. We tested it with specifically designed Procellariiformes-targeted primer sets for rapid sexing of the birds using fluorimetric detection. The protocol, combined with rapid DNA extraction, allows for fast reading of results without electrophoresis within less than 1 h from sampling. The technique could be extended to other species, as well as to many other applications. |
| first_indexed | 2024-03-11T09:07:51Z |
| format | Article |
| id | doaj.art-352f8cd55f7e4b1d8af2f2b712b795ff |
| institution | Directory Open Access Journal |
| issn | 2079-7737 |
| language | English |
| last_indexed | 2024-03-11T09:07:51Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biology |
| spelling | doaj.art-352f8cd55f7e4b1d8af2f2b712b795ff2023-11-16T19:14:30ZengMDPI AGBiology2079-77372023-02-0112230010.3390/biology12020300High-Performance PCR for Alleles Discrimination of Chromo-Helicase-DNA Binding Protein (CHD1) Gene in Bird SexingMarcello Tagliavia0Valentina Catania1Giacomo Dell’Omo2Bruno Massa3Institute for Biomedical Research and Innovation-National Research Council (IRIB-CNR), Via Ugo La Malfa 153, 90146 Palermo, ItalyDepartment of Earth and Marine Sciences (DiSTeM), University of Palermo, Viale delle Scienze, bd. 16, 90128 Palermo, ItalyOrnis Italica, Piazza Crati 15, 00199 Roma, ItalyDepartment of Agriculture, Food and Forest Sciences (SAAF), University of Palermo, Viale delle Scienze, bd. 13, 90128 Palermo, ItalyGenetic analyses aiming at assessing the presence of specific sequences or alleles are often carried out by PCR. Sexing of most birds is nowadays based on PCR with “universal” primers and relies on the assessment of the presence of the sex-linked CHD1-Z and -W alleles. The entire workflow is relatively time-consuming, especially for batch analyses, whereas methods that allow carrying out the entire procedure in a short time are highly desirable. The only method for outdoor analyses reported so far relies on LAMP; however; it fails to work properly in Procellariiformes. Besides improving the LAMP test; we have developed a PCR-based DNA amplification procedure (named high-performance PCR); whose unique features allow it to outperform standard PCR; making possible the direct, in-tube visual reading of results. We tested it with specifically designed Procellariiformes-targeted primer sets for rapid sexing of the birds using fluorimetric detection. The protocol, combined with rapid DNA extraction, allows for fast reading of results without electrophoresis within less than 1 h from sampling. The technique could be extended to other species, as well as to many other applications.https://www.mdpi.com/2079-7737/12/2/300high-performance PCRloop-mediated isothermal amplification (LAMP)fast DNA extractionrapid bird sexingProcellariiformes |
| spellingShingle | Marcello Tagliavia Valentina Catania Giacomo Dell’Omo Bruno Massa High-Performance PCR for Alleles Discrimination of Chromo-Helicase-DNA Binding Protein (CHD1) Gene in Bird Sexing Biology high-performance PCR loop-mediated isothermal amplification (LAMP) fast DNA extraction rapid bird sexing Procellariiformes |
| title | High-Performance PCR for Alleles Discrimination of Chromo-Helicase-DNA Binding Protein (CHD1) Gene in Bird Sexing |
| title_full | High-Performance PCR for Alleles Discrimination of Chromo-Helicase-DNA Binding Protein (CHD1) Gene in Bird Sexing |
| title_fullStr | High-Performance PCR for Alleles Discrimination of Chromo-Helicase-DNA Binding Protein (CHD1) Gene in Bird Sexing |
| title_full_unstemmed | High-Performance PCR for Alleles Discrimination of Chromo-Helicase-DNA Binding Protein (CHD1) Gene in Bird Sexing |
| title_short | High-Performance PCR for Alleles Discrimination of Chromo-Helicase-DNA Binding Protein (CHD1) Gene in Bird Sexing |
| title_sort | high performance pcr for alleles discrimination of chromo helicase dna binding protein chd1 gene in bird sexing |
| topic | high-performance PCR loop-mediated isothermal amplification (LAMP) fast DNA extraction rapid bird sexing Procellariiformes |
| url | https://www.mdpi.com/2079-7737/12/2/300 |
| work_keys_str_mv | AT marcellotagliavia highperformancepcrforallelesdiscriminationofchromohelicasednabindingproteinchd1geneinbirdsexing AT valentinacatania highperformancepcrforallelesdiscriminationofchromohelicasednabindingproteinchd1geneinbirdsexing AT giacomodellomo highperformancepcrforallelesdiscriminationofchromohelicasednabindingproteinchd1geneinbirdsexing AT brunomassa highperformancepcrforallelesdiscriminationofchromohelicasednabindingproteinchd1geneinbirdsexing |