KHẢO SÁT QUY TRÌNH LY TRÍCH NHANH DNA - ĐỀ XUẤT QUY TRÌNH LY TRÍCH TỐI ƯU CHO NIÊM MẠC MIỆNG ỨNG DỤNG TRONG XÉT NGHIỆM QUAN HỆ HUYẾT THỐNG

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Ngôn ngữ: vie

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Thông tin xuất bản: Tạp chí Khoa học và Công nghệ - Đại học Thái Nguyên, 2020

Mô tả vật lý: tr.45726

Bộ sưu tập: Báo, Tạp chí

ID: 303282

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After experiment, the process used NaOH - SDS combinate with thermal shock was found to be the DNA extraction method more efficient than the other. Comparing extraction efficiency at different SDS concentrations to propose the optimal extraction procedure, the result showed that 0.01% is the suitable SDS concentration. To evaluate the effectiveness of the improved extracting procedure, we compared it with a commercially available kit (E.Z.N.A Forensic DNA kit) on 10 samples and applicated to extract DNA on 200 samples. After extracted, all samples have been used for STR-PCR reaction. On 10 samples, average fluorescence signal from the improved extraction procedure is lower than the commercial kit, but the results were accurate and consistent. On 200 samples, the results showed that no samples were inhibited, more than 90% of samples for PCR product had fluorescence intensity above 100 RFU and 100% for signal above 50 RFU. On the samples have been known the parentage relationship, the result is match.Khảo sát một số quy trình ly trích đã có cho thấy, phương pháp ly trích nhanh DNA bằng NaOH -SDS kết hợp với sốc nhiệt cho kết quả khả quan hơn so với các phương pháp còn lại. Khi so sánh hiệu quả ly trích ở các nồng độ SDS khác nhau, 0,01% là nồng độ SDS tối ưu. Quy trình đề xuất được đánh giá thông qua việc so sánh với bộ ly trích cột thương mại OMEGA và ly trích thử nghiệm trên 200 mẫu niêm mạc miệng. Kết quả so sánh với bộ ly trích thương mại, DNA ly trích bởi quy trình đề xuất cho sản phẩm PCR có cường độ huỳnh quang trung bình thấp hơn nhưng kết quả phân tích vẫn chính xác và trùng khớp. Trên 200 mẫu thực tế, không có mẫu nào bị ức chế, hơn 90% mẫu cho sản phẩm PCR có cường độ huỳnh quang trên 100 RFU và 100% cho tín hiệu cao trên 50 RFU. Đối với các cặp mẫu đã biết mối quan hệ huyết thống, kết quả trả về là trùng khớp.

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