DNA Isolation from Cocoa-Derived Products and Cocoa Authentication by TaqMan Real-Time PCR

• de Oliveira, A. C. , Muhovski, Y. , Rogez, H. & Debode, F. (2023). DNA Isolation from Cocoa-Derived Products and Cocoa Authentication by TaqMan Real-Time PCR. PCR: Methods and Protocols, 2967: 75-83.

Type	Journal Article
Year	2023
Title	DNA Isolation from Cocoa-Derived Products and Cocoa Authentication by TaqMan Real-Time PCR
Journal	PCR: Methods and Protocols
Label	U1-Muhovski
Volume	2967
Pages	75-83
Date	october
Endnote Keywords	Chloroplast DNA, Chocolate, DNA extraction, Nuclear DNA, Theobroma cacao, qPCR
Abstract	Cocoa (Theobroma cacao L.) is an international commodity used as an ingredient in the manufacturing of chocolate making its authentication a key issue in the cocoa chain. Various molecular techniques have been increasingly applied for quality requirements. These issues highlight the need for techniques that allow the extraction and detection of cocoa DNA from highly processed cocoa products and chocolate. The applicability of real-time PCR to highly processed cocoa-derived products for authentication purposes depends on the possibility of extracting high-quality and amplifiable DNA and further developing efficient PCR tests. This methodology herein describes the use of a classical CTAB method providing DNA suitable for TaqMan real-time PCR amplification. Real-time PCR is a simple and fast method, with a high potential application in a wide range of food products. The main features of this technique are focused on two DNA targets, one located in the nuclear genome (vicilin-li PCR test) and a second one based on chloroplast DNA (lipids PCR test), which successfully passed the performance criteria considering the specificity, sensitivity, efficiency of amplification, robustness, and applicability in processed cocoa-derived products and chocolate.
Fichier
Lien	https://link.springer.com/protocol/10.1007/978-1-0716-3358-8_7
Authors	de Oliveira, A. C., Muhovski, Y., Rogez, H., Debode, F.