Lâm Thị Việt Hà * , Hà Thanh Toàn , Trần Nhân Dũng Trương Trọng Ngôn

* Tác giả liên hệ (ltvha@ctu.edu.vn)

Abstract

Four methods for extracting DNA in cocoa-derived products (cocoa powder, cocoa butter, cocoa mass, and dark chocolate) are described. These DNA extraction methods included four commercial kits DNeasy Plant kit, DNeasy Plant kit with polyvinyl polypyrrolidone (PVPP), QIAamp DNA Stool kit, Wizard Genomic DNA Purification kit. The DNA yield, purity, and quality for four different cocoa matrices are discussed. The yield and purity were determined through spectrophotometry: DNA concentration, DNA purity and PCR positive amplification. Three different conventional PCR reactions were used to assess the quality of the DNA extracted. The Wizard Genomic DNA Purification kit was found to be the best for DNA quality and PCR detection in cocoa powder and dark chocolate ([DNA] = 44.83 (ng/µL); A260/A280 = 1.4-1.7, PCR positive amplification) and ([DNA] = 29.34 (ng/µL) and A260/A280 = 1.4-1.7, PCR positive result, respectively). The cocoa butter matrix gave the lowest DNA yield, and negative amplification results for three the examined extraction methods DNeasy Plant kit with PVPP, DNeasy Plant kit without PVPP, and QIAamp DNA Stool kit [DNA] <10 (ng/µL, PCR negative products). The QIAamp DNA Stool kit method showed the best result for PCR detection of cocoa mass (A260/A280 = 1.7-2.0, [DNA] = 28.12 (ng/µL), PCR negative amplification).
Keywords: Cocoa, cocoa-derived product, DNA extraction

Tóm tắt

Bốn phương pháp trích ly DNA nhanh (kit) được dùng trong nghiên cứu đánh giá và so sánh nhằm chọn phương pháp trích ly DNA tối ưu nhất cho từng nguyên liệu có nguồn gốc từ sản phẩm ca cao (bốn nguyên liệu sử dụng trong thí nghiệm: bột ca cao, bơ ca cao, ca cao lỏng và sô cô la). Các phương pháp trích ly bao gồm: DNeasy Plant kit, DNeasy Plant kit with polyvinyl polypyrrolidone (PVPP), QIAamp DNA Stool kit, và Wizard Genomic DNA Purification kit. DNA ly trích được so sánh nồng độ DNA; độ tinh sạch và so sánh chất lượng sản phẩm tạo thành (sản phẩm PCR dương tính); Wizard Genomic DNA Purification kit cho kết quả tốt nhất đối với bột ca cao và sô cô la [DNA] = 44,83 (ng/µL); A260/A280 = 1,4-1,7, PCR dương tính; và [DNA] = 29,34 (ng/µL) và A260/A280 = 1,4-1,7, PCR dương tính; sản phẩm bơ ca cao không cho kết quả tốt đối với 3 phương pháp ly trích DNeasy Plant kit with PVPP, DNeasy Plant kit without PVPP, và QIAamp DNA Stool kit ([DNA] <10 (ng/µL PCR âm tính); QIAamp DNA Stool kit cho kết quả tối ưu để ly trích DNA từ ca cao lỏng (A260/A280 = 1,7-2.0, ([DNA] = 28,12 (ng/µL), PCR dương tính.
Từ khóa: Ca cao, PCR, phương pháp trích ly DNA

Article Details

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