Meat species identification by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of mitochondrial 12S rRNA gene

Girish, P. S.; Anjaneyulu, A. S. R.; Viswas, K.N.; Shivakumar, B. M.; Anand, Manish Kumar; Patel, Mukesh R.; Sharma, Bhaskar

doi:10.1016/j.meatsci.2004.12.004

Cited by 168 publications

(102 citation statements)

References 18 publications

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“…Also, AL method does not require any centrifugation unlike that of commercial kit and PCI based methods. Chances of contamination of samples is also less as it is a single tube method which avoids contamination during pipetting and transferring the sample to other tubes PCR amplification using the DNA extracted by different methods For checking the suitability of DNA extracted from meat, heart, liver & kidney extract by AL method, extract was subjected to PCR amplification using universal primers of mitochondrial 12S rRNA gene (Girish et al 2005). PCR amplification yielded amplicon of size of about 456 bp and the result of PCR amplification of mitochondrial 12S rRNA gene is given in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Test was repeated in ten different buffalo meat samples. Girish et al (2005) utilized PCR RFLP technique for identification of buffalo meat which involved PCR amplification of mitochondrial 12S rRNA gene (amplicon size 456 bp) followed by restriction digestion with HhaI restriction enzyme targeting species specific restriction site, which yielded products of size 247 bp and 209 bp after digestion. A buffalo meat certification method based on PCR amplification of mitochondrial Cytochrome B gene followed by restriction digestion with TaqI restriction enzyme yielding digestion products of 108 bp and 163 bp was reported by Teixeira et al (2007).…”

Section: Resultsmentioning

confidence: 99%

“…PCR amplification of Mitochondrial 12S rRNA gene Universal primers for mt 12S rRNA gene used for speciation of meat (Girish et al 2005) was used and sequence of primers are as given below:…”

Section: Methodsmentioning

confidence: 99%

“…The DNA molecule is more stable and the composition of DNA is same in any cell of the individual. Hence, DNA based techniques namely Polymerase Chain Reaction (PCR) and its variants: Restriction Fragment Length Polymorphism (RFLP) (Girish et al 2005); Random Amplified Polymorphic DNA (RAPD) finger printing (Saez et al 2004); DNA Hybridization (Buntjer et al 1999), PCR Sequencing (Girish et al 2007); Arbitrarily Primed-PCR (AP-PCR) (Desmarais et al 1998) and PCR-Single Strand Conformation Polymorphisms (SSCP) (Tejedor et al 2006) and species specific markers (Haunshi et al 2009) have been employed for the identification of various animal species in the recent past. The PCR-based techniques provide high level of specificity, sensitivity, accuracy and precision than the techniques hitherto being used for the purpose of animal species identification.…”

Section: Introductionmentioning

confidence: 99%

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A rapid method for authentication of Buffalo (Bubalus bubalis) meat by Alkaline Lysis method of DNA extraction and species specific polymerase chain reaction

et al. 2011

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Buffalo (Bubalus bubalis) meat is a major item of export from India but export of beef i.e. meat from cattle (Bos indicus) is prohibited. Also, adulteration of buffalo meat with that of beef (meat from cattle) is a common fraudulent practice because of prohibition on cow slaughter in most states of India. Food analysts require precise identification techniques to implement such regulations. In the present study, a method of DNA extraction by Alkaline lysis from meat samples and speciation of buffalo meat using species specific Polymerase Chain Reaction (PCR) has been reported. Alkaline lysis technique is a rapid method which involves triturating meat with four volumes of 0.2N NaOH, dilution of resultant liquid extract with eight volumes of 0.2N NaOH, heating the mix 75°C for 20 min followed by neutralization with eight volumes of 0.04N Tris HCl. Entire procedure of DNA extraction takes less than 30 min and it is economical as it involves less expensive chemicals. Method was successfully applied in animal byproducts also viz., liver, heart and kidney. For authentication of buffalo meat, pair of primers was designed based on mitochondrial D loop gene nucleotide sequence. PCR amplification using the designed primers gave amplicon of size 482 bp in buffalo and no amplification was detected in closely related species viz., cattle, sheep and goat meat samples. Results of the assay were highly repetitive and reliable. An export sample referred by export regulation authorities was also analyzed by using the Alkaline lysis method of DNA extraction and species specific PCR which enabled authentication of meat within 5 h.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A rapid method for authentication of Buffalo (Bubalus bubalis) meat by Alkaline Lysis method of DNA extraction and species specific polymerase chain reaction

et al. 2011

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“…Although molecular-based techniques are not the only tools used for species identification, even recently, advances in molecular techniques over the last two decades have allowed the development of authentic and reliable methods for species identification (Nijman et al, 2003;Girish et al, 2005). These techniques are promising and are able to overcome the drawbacks of many conventional methods.…”

Section: Introductionmentioning

confidence: 99%

Species identification of Malayan Gaur, Kedah-Kelantan and Bali cattle using polymerase chain reaction-restricted fragment length polymorphism

et al. 2014

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ABSTRACT. Mitochondrial DNA (mtDNA) is a useful genetic marker that can be used for species identification. The cytochrome b (Cyt b) gene is a suitable mtDNA candidate gene for use in phylogenetic analyses due to its sequence variability, which makes it appropriate for comparisons at the subspecies, species, and genus levels. This study was conducted to develop a rapid molecular method for species identification of Malayan gaur (Bos gaurus hubbacki), Kedah-Kelantan (KK) (Bos indicus), and Bali (Bos javanicus) cattle in Malaysia. DNA was extracted from blood samples of 8 Malayan gaurs, 30 KK, and 28 Bali cattle. A set of both specific and universal primers for the Cyt b gene were used in PCR amplification. DNA sequences obtained were then analyzed using BioEdit and Restriction Mapper softwares. The PCR products obtained from Cyt b gene amplification were then subjected to restriction enzyme Species identification of Malaysian cattle with PCR-RFLP digestion. The amplification, using both specific and universal primers, produced a 154-and a 603-bp fragment, respectively, in all three species. Two restriction enzymes, NlaIV and SspI, were used to obtain specific restriction profiles that allowed direct identification of Malayan gaur, KK, and Bali cattle. Our findings indicate that all three species can be identified separately using a combination of universal primers and the restriction enzyme SspI.

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Detecting Adulteration in Halal Foods

Farag

2020

The Halal Food Handbook

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Meat species identification by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of mitochondrial 12S rRNA gene

Cited by 168 publications

References 18 publications

A rapid method for authentication of Buffalo (Bubalus bubalis) meat by Alkaline Lysis method of DNA extraction and species specific polymerase chain reaction

A rapid method for authentication of Buffalo (Bubalus bubalis) meat by Alkaline Lysis method of DNA extraction and species specific polymerase chain reaction

Species identification of Malayan Gaur, Kedah-Kelantan and Bali cattle using polymerase chain reaction-restricted fragment length polymorphism

Detecting Adulteration in Halal Foods

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