GB/T 35918-2018 in English

1. Background and significance of the standard

GB/T 35918-2018 "Sanger sequencing method for detecting animal origin in animal products" is an important standard for the identification of the origin of animal products. It is mainly applicable to the qualitative detection of meat, milk, blood, hair, horn, bone, viscera, hides and other products made from mammals, poultry, fish and other animals.

2. Technical principles and methods

4. Sample processing and detection process

4.1 Sample collection and preparation

According to different sample types, use the corresponding sampling method (GB/T 9695.19, GB/T 14699.1 and SN/T 3500-2013) for processing, and store as required.

4.2 DNA extraction and purification

Use CTAB method or equivalent kit to extract DNA to ensure that the DNA concentration and purity meet the detection requirements (OD260/OD280 ratio should be between 1.7-2.1).

4.3 PCR amplification and electrophoresis detection

Select appropriate primers according to the target species, perform PCR amplification, and detect the quality of the amplified product by agarose gel electrophoresis.

4.4 Sequencing and data analysis

Use Sanger sequencer to perform bidirectional sequencing on the PCR product, remove the low-quality region by splicing software, and then compare and confirm the species in the BOLD or NCBI database.