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A Novel Direct PCR Lysis Buffer Can Improve PCR from Meat Matrices

Author:
Guan, Feng, Jin, Yuting, Zhao, Jin, Ai, Juntao, Luo, Yuanyuan
Source:
Food analytical methods 2019 v.12 no.1 pp. 100-107
ISSN:
1936-9751
Subject:
DNA, DNA barcoding, DNA replication, EDTA (chelating agent), absorbance, analytical methods, cost effectiveness, genetic analysis, liquids, polymerase chain reaction, polysorbates, processed meat, sodium hydroxide
Abstract:
Molecular technologies based on PCR have been widely used in many biological analysis fields, especially for genetic analysis and DNA barcoding. In this study, a rapid DNA lysis liquid was formulated without any purification step from fresh and processed meat, suitable for conventional PCR amplification. Three different lysis liquid formulas were designed for selection and further optimization for direct PCR and absorbance spectra; DNA concentration and performance in PCR were used to assess the effect of each formula. The results indicated that the formula containing NaOH, EDTA, SDS, Tween 20, and Tris-HCl achieved the best results, and the optimized formula met the need of practical PCR applications. The protocol provided a rapid lysis buffer for DNA replication from any meat samples. The performance of the final formula resulted in high DNA lysis efficiencies for all the tested meat samples and the PCR amplification efficiencies were similar to isolated DNA template using a commercial kit. The whole process can be completed in 30 min. Therefore, this study provides a simple, alternative, cost-effective fast solution for meat molecular analysis based on DNA.
Agid:
6275861