Research Article
GDO ANALİZLERİNDE KULLANILAN CRM İZOLATLARININ KONTROLLÜ MUHAFAZA KOŞULLARINDA TEKRAR KULLANIM OLANAKLARININ ARAŞTIRILMASI
Abstract
Genetiği değiştirilmiş organizmalar (GDO) potansiyel yararları nedeniyle tarımsal ürünler başta olmak üzere birçok alanda kullanılmaktadır. Yasal düzenlemelere göre gıda ve yemlerde GDO varlığının bilinmesi zorunluluktur. Analizlerde sonuçların doğruluğu, kalibrasyon eğrisinin hazırlanması ve kalite kontrol çalışmalarında sertifikalı referans maddeler (CRM) kullanılmaktadır. Çalışma ile bitkisel ürünlerde GDO analizlerinde kullanılan CRM izolatlarının tekrar kullanılabilme olanaklarının belirlenmesi amacıyla dondur-çözdür uygulaması ile meydana gelebilecek degradasyon araştırılmıştır. Bunun için soya ve mısır CRM’lerinden elde edilen izolatlara kontrollü koşullarda belirli periyotlarla (5, 10, 15, 20 kez) dondurma (-20±2°C) ve çözdürme (22±2°C) işlemleri uygulanmıştır. Real time PCR cihazında yapılan analizler sonucunda 20 dondur-çözdür uygulamasına kadar CRM izolatlarının tekrar kullanılabildiği görülmüştür.
Keywords
Supporting Institution
Gıda ve Yem Kontrol Merkez Araştırma Enstitüsü Müdürlüğü
Project Number
TAGEM/GYKMAE/A/19/A3/P7/01
Thanks
Bu çalışmayı TAGEM/GYKMAE/A/19/A3/P7/01 proje numarası ile maddi olarak destekleyen Tarımsal Araştırmalar ve Politikalar Genel Müdürlüğü’ne teşekkür ederiz.
References
- Al-Warid, H. S. (2014). The Effect Of Freezıng-Thawıng On DNA Extractıon From Cryptosporıdıum Oocysts In Fecal Samples. https://www.ijpbs.com/ijpbsadmin/upload/ijpbs_52f98de2e3214.pdf
- Bellete, B., Flori, P., Hafid, J., Raberin, H., Tran Manh Sung, R. (2003). Influence of the quantity of nonspecific DNA and repeated freezing and thawing of samples on the quantification of DNA by the Light Cycler, Journal of Microbiological Methods, 55, 213–219.
- Brunstein, J., (2015). Freeze-thaw cycles and nucleic acid stability: what’s safe for your samples?. https://www.mlo-online.com/freeze-thaw-cycles-and-nucleic-acid-stability-whats-safe-for-your-samples.php
- Bus´, M.M., Allen, M. (2014). Collecting and Preserving Biological Samples from Challenging Environments for DNA Analysis, Biopreservation and biobanking, 12(1), 17-22.
- Commission Regulation (EC) No 1829/2003 of the European Parliament and Council of 22 September 2003 on genetically modified food and feed. (2003). Official Journal of the European Union, L 268/1, 18 October 2003.
- Eurofins GENESpin (2016). Kit for isolation of high-quality DNA from food and feed samples.
- European Commission (2009). Event-specific Method for the Quantification of Soybean Event DP-305423-1 Using Real-time. Joint Research Centre Institute for Health and Consumer Protection Molecular Biology and Genomics Unit.
- European Commission (2012). Event-specific method for the quantification of maize line DAS40278-9 using real-time PCR Protocol. Joint Research Centre Institute for Health and Consumer Protection Molecular Biology and Genomics Unit.
- European Commission (2013). Event-specific Method for the Quantification of Soybean Event DP-305423-1 Using Real-time PCR v. 1.01. Joint Research Centre Institute for Health and Consumer Protection Molecular Biology and Genomics Unit.
- EU SCIENCE HUB. The European Commission’s science and knowledge service. Reference materials for GMO analysis. https://ec.europa.eu/jrc/en/research-topic/reference-materials-gmo-analysis.
- European Union. (2017). Recommendation for the unit of measurement and the measuring system to report traceable and comparable results expressing GM content in accordance with EU legislation (JRC106032 EUR 28536 EN). Luxembourg: Publications Office of the European Union
- Fan, X. J., Huang, Y., Wu, P. H., Yin, X. K., Yu, X. H., Fu, X. H., ... ve Yin, J. X. (2019). Impact of cold ischemic time and freeze-thaw cycles on RNA, DNA and protein quality in colorectal cancer tissues biobanking, Journal of Cancer, 10(20), 4978.
- Gaikwad, A.B. (2020). DNA storage methodologies: Principles and Protocols. http:// www.nbpgr.ernet.in/Portals/6/DMX/GENOMIC_RESOURCES/DNA%20storage%20methods-principles%20and%20protocols.pdf
- GDO Analizlerinde Verifikasyon Rehberi. (2018). T.C. Tarım ve Orman Bakanlığı Gıda ve Kontrol Genel Müdürlüğü, Nisan 2018.
- Lahiri, D. K., Schnabel, B. (1993). DNA isolation by a rapid method from human blood samples: effects of MgCl2, EDTA, storage time, and temperature on DNA yield and quality, Biochemical genetics, 31(7-8), 321-328.
- Lee, S. B., Crouse, C. A., Kline, M. C. (2010). Optimizing storage and handling of DNA extracts. http://books.google.com/books (Original work published 2010)
- Rice, B. J., Haussler, D. (2018). U.S. Patent Application No. 15/561,447. Google Patents.
- Ross, K. S., Haites, N. E., Kelly, K. F. (1990). Repeated freezing and thawing of peripheral blood and DNA in suspension: effects on DNA yield and integrity, Journal of medical genetics, 27(9), 569-570.
- Rossmanith, P., Röder, B., Frühwirth, K., Vogl, C., Wagner, M. (2011). Mechanisms of degradation of DNA standards for calibration function during storage, Applied microbiology and biotechnology, 89(2), 407-417.
- Röder, B., Frühwirth, K., Vogl, C., Wagner, M., Rossmanith, P. (2010). Impact of Long-Term Storage on Stability of Standard DNA for Nucleic Acid-Based Methods, Journal Of Clinical Microbiology, 48(11), 4260–4262
- Sanlidag, T., Akcali, S., Ozbakkaloglu, B. (2005). Serum hepatitis B DNA: stability in relation to multiple freeze–thaw procedures, Journal of virological methods, 123 (1), 49-52.
- Schaudien, D., Baumgärtner, W., Herden, C. (2007). High preservation of DNA standards diluted in 50% glycerol, Diagnostic molecular pathology, 16(3), 153-157.
- Shao, W., Khin, S., Kopp, W. C. (2012). Characterization of effect of repeated freeze and thaw cycles on stability of genomic DNA using pulsed field gel electrophoresis, Biopreservation and biobanking, 10(1), 4-11.
- Ünal, T.T. (2019). Gıda Ve Yem Ürünlerinde GDO Tespiti İçin Yeterlilik Test Kiti Geliştirilmesi. (Yüksek lisans tezi). Hacettepe Üniversitesi, Fen Bilimleri Enstitüsü, Ankara.
- Widell, A., Månsson, A. S., Sundstrom, G., Hansson, B. G., Nordenfelt, E. (1991). Hepatitis C virus RNA in blood donor sera detected by the polymerase chain reaction: comparison with supplementary hepatitis C antibody assays, Journal of medical virology, 35(4), 253-258.
INVESTIGATION OF REUSE CAPABILITIES OF CRM ISOLATES USED IN GMO ANALYSIS IN THE CONTROLLED STORAGE CONDITIONS
Abstract
Genetically modified organisms (GMOs) are used in many areas, especially in agricultural crops, due to their potential benefits. According to legal regulations, it is imperative to know the presence of GMOs in food and feed. Certified reference materials (CRM) are used in the accuracy of the analyzes, preparation of calibration curve, and in quality control studies. In this study, degradation occuring with freeze-thaw application has been investigated in order to determine the reuse possibilities of CRM isolates used in GMO analysis in herbal products. For this, freezing (-20 ± 2 ° C) and thawing (22 ± 2 ° C) processes were applied to the isolates obtained from soy and corn CRMs at certain periods (5, 10, 15, 20 times) under controlled conditions. Real time PCR analysis results showed that up to 20 freeze-thaw applications, CRM isolates were reusable.
Keywords
Project Number
TAGEM/GYKMAE/A/19/A3/P7/01
References
- Al-Warid, H. S. (2014). The Effect Of Freezıng-Thawıng On DNA Extractıon From Cryptosporıdıum Oocysts In Fecal Samples. https://www.ijpbs.com/ijpbsadmin/upload/ijpbs_52f98de2e3214.pdf
- Bellete, B., Flori, P., Hafid, J., Raberin, H., Tran Manh Sung, R. (2003). Influence of the quantity of nonspecific DNA and repeated freezing and thawing of samples on the quantification of DNA by the Light Cycler, Journal of Microbiological Methods, 55, 213–219.
- Brunstein, J., (2015). Freeze-thaw cycles and nucleic acid stability: what’s safe for your samples?. https://www.mlo-online.com/freeze-thaw-cycles-and-nucleic-acid-stability-whats-safe-for-your-samples.php
- Bus´, M.M., Allen, M. (2014). Collecting and Preserving Biological Samples from Challenging Environments for DNA Analysis, Biopreservation and biobanking, 12(1), 17-22.
- Commission Regulation (EC) No 1829/2003 of the European Parliament and Council of 22 September 2003 on genetically modified food and feed. (2003). Official Journal of the European Union, L 268/1, 18 October 2003.
- Eurofins GENESpin (2016). Kit for isolation of high-quality DNA from food and feed samples.
- European Commission (2009). Event-specific Method for the Quantification of Soybean Event DP-305423-1 Using Real-time. Joint Research Centre Institute for Health and Consumer Protection Molecular Biology and Genomics Unit.
- European Commission (2012). Event-specific method for the quantification of maize line DAS40278-9 using real-time PCR Protocol. Joint Research Centre Institute for Health and Consumer Protection Molecular Biology and Genomics Unit.
- European Commission (2013). Event-specific Method for the Quantification of Soybean Event DP-305423-1 Using Real-time PCR v. 1.01. Joint Research Centre Institute for Health and Consumer Protection Molecular Biology and Genomics Unit.
- EU SCIENCE HUB. The European Commission’s science and knowledge service. Reference materials for GMO analysis. https://ec.europa.eu/jrc/en/research-topic/reference-materials-gmo-analysis.
- European Union. (2017). Recommendation for the unit of measurement and the measuring system to report traceable and comparable results expressing GM content in accordance with EU legislation (JRC106032 EUR 28536 EN). Luxembourg: Publications Office of the European Union
- Fan, X. J., Huang, Y., Wu, P. H., Yin, X. K., Yu, X. H., Fu, X. H., ... ve Yin, J. X. (2019). Impact of cold ischemic time and freeze-thaw cycles on RNA, DNA and protein quality in colorectal cancer tissues biobanking, Journal of Cancer, 10(20), 4978.
- Gaikwad, A.B. (2020). DNA storage methodologies: Principles and Protocols. http:// www.nbpgr.ernet.in/Portals/6/DMX/GENOMIC_RESOURCES/DNA%20storage%20methods-principles%20and%20protocols.pdf
- GDO Analizlerinde Verifikasyon Rehberi. (2018). T.C. Tarım ve Orman Bakanlığı Gıda ve Kontrol Genel Müdürlüğü, Nisan 2018.
- Lahiri, D. K., Schnabel, B. (1993). DNA isolation by a rapid method from human blood samples: effects of MgCl2, EDTA, storage time, and temperature on DNA yield and quality, Biochemical genetics, 31(7-8), 321-328.
- Lee, S. B., Crouse, C. A., Kline, M. C. (2010). Optimizing storage and handling of DNA extracts. http://books.google.com/books (Original work published 2010)
- Rice, B. J., Haussler, D. (2018). U.S. Patent Application No. 15/561,447. Google Patents.
- Ross, K. S., Haites, N. E., Kelly, K. F. (1990). Repeated freezing and thawing of peripheral blood and DNA in suspension: effects on DNA yield and integrity, Journal of medical genetics, 27(9), 569-570.
- Rossmanith, P., Röder, B., Frühwirth, K., Vogl, C., Wagner, M. (2011). Mechanisms of degradation of DNA standards for calibration function during storage, Applied microbiology and biotechnology, 89(2), 407-417.
- Röder, B., Frühwirth, K., Vogl, C., Wagner, M., Rossmanith, P. (2010). Impact of Long-Term Storage on Stability of Standard DNA for Nucleic Acid-Based Methods, Journal Of Clinical Microbiology, 48(11), 4260–4262
- Sanlidag, T., Akcali, S., Ozbakkaloglu, B. (2005). Serum hepatitis B DNA: stability in relation to multiple freeze–thaw procedures, Journal of virological methods, 123 (1), 49-52.
- Schaudien, D., Baumgärtner, W., Herden, C. (2007). High preservation of DNA standards diluted in 50% glycerol, Diagnostic molecular pathology, 16(3), 153-157.
- Shao, W., Khin, S., Kopp, W. C. (2012). Characterization of effect of repeated freeze and thaw cycles on stability of genomic DNA using pulsed field gel electrophoresis, Biopreservation and biobanking, 10(1), 4-11.
- Ünal, T.T. (2019). Gıda Ve Yem Ürünlerinde GDO Tespiti İçin Yeterlilik Test Kiti Geliştirilmesi. (Yüksek lisans tezi). Hacettepe Üniversitesi, Fen Bilimleri Enstitüsü, Ankara.
- Widell, A., Månsson, A. S., Sundstrom, G., Hansson, B. G., Nordenfelt, E. (1991). Hepatitis C virus RNA in blood donor sera detected by the polymerase chain reaction: comparison with supplementary hepatitis C antibody assays, Journal of medical virology, 35(4), 253-258.
There are 25 citations in total.
Details
| Primary Language | Turkish |
|---|---|
| Subjects | Food Engineering |
| Journal Section | Articles |
| Authors | |
| Project Number | TAGEM/GYKMAE/A/19/A3/P7/01 |
| Publication Date | May 15, 2021 |
| Published in Issue | Year 2021 Volume: 46 Issue: 3 |
