Primers and PCR conditions used in this study
| Exons | Primers | Sequence | PCR conditions | PCR product |
|---|---|---|---|---|
| 1 | 1-forward | 5'-GGGTGTGGCTGATTTACATCC-3' | 94°C for 2 min, followed by 31 cycles (94°C for 30 s, 62°C for 30 s,72°C for 30 s), and a final elongation at 72°C for 5 min | 340 bp |
| 1-reverse | 5'-CCAGAGGCATCCAGACTCG-3' | |||
| 2 | 2-forward | 5'-AAAGGTTGTCTGTGCGAATGT-3' | 94°C for 2 min, followed by 31 cycles (94°C for 30 s, 64°C for 30 s,72°C for 30 s), and a final elongation at 72°C for 5 min | 319 bp |
| 2-reverse | 5'-CCAGAGGGTGCTCTAGGGAG-3' | |||
| 3 | 3-forward | 5'-ATCAAAATGCAATGCAGCTAAA-3' | 94°C for 2 min, followed by 31 cycles (94°C for 30 s, 62°C for 30 s,72°C for 30 s), and a final elongation at 72°C for 5 min | 635 bp |
| 3-reverse | 5'-AGACTTCTGGAAATGTTGCCTA-3' | |||
| 4 | 4-forward | 5'-GGGGGAGTGGATCAAC-3' | 94°C for 2 min, followed by 36 cycles (94°C for 30 s, 60°C for 30 s,72°C for 30 s), and a final elongation at 72°C for 5 min | 540 bp |
| 4-reverse | 5'-GTCTGCCAGGATGTCTTAACC-3' | |||
| 5 | 5-forward | 5'-TGGCAGACAGATAACT-3' | 94°C for 2 min, followed by 32 cycles (94°C for 30 s, 55°C for 30 s,72°C for 30 s), and a final elongation at 72°C for 5 min | 717 bp |
| 5-reverse | 5'-CTCTCTCCTCATATCTCTC-3' |
Study conception and design were performed within the context of the postgraduate program “Clinical Applications of Molecular Medicine”, Faculty of Medicine, School of Health Sciences, University of Thessaly, Greece. The authors express their sincere gratitude to Lemonia Anagnostopoulos for her invaluable assistance in providing important comments on the quality of the manuscript.
ML: Conceptualization, Investigation, Data curation, Formal analysis, Software, Methodology, Writing—original draft, Writing—review & editing. SS: Validation, Writing—review & editing, Software, Methodology, Writing—original draft, Writing—review & editing. MS: Conceptualization, Writing—original draft, Writing—review & editing, Supervision. NG, EH, HP, AD, VA, AM, and VK: Investigation, Validation, Writing—review & editing. GV, CH, and FK: Validation, Writing—review & editing. All authors read and approved the submitted version.
The authors declare that they have no conflicts of interest.
The study was approved by the institutional review boards of the University Hospital of Larissa, Greece (14/17/29.11.2022) and the Faculty of Medicine of the University of Thessaly, Greece (4/16.01.2023).
Written informed consent was obtained from each individual or an accompanying relative, in the case of patients where consent was not legally applicable (e.g., children).
Not applicable.
The raw data supporting the conclusions of this manuscript will be made available by the authors, without undue reservation, to any qualified researcher.
The study was supported by the standard budget provided by The Research Committee of the University of Thessaly to Department of Immunology & Histocompatibility, Faculty of Medicine, University of Thessaly, 41500 Larissa, Greece. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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