Abstract
We describe a study on the use of rolling circle amplification (RCA) for detecting GM event-specific motifs within short PCR amplicons, synthetic oligonucleotides, and extracted plant genomic DNA targets, as an alternative to the polymerase chain reaction (PCR). PCR-based detection has limitations that include the cost of reagents and equipment, and the potential for erroneous amplification of a contaminant. Our results reveal that RCA enables discrimination between the wild type (wt) and GM motifs when the sequences are within short PCR amplicons or synthetic oligonucleotides, but not within plant genomic DNA. These findings highlight the potential problem with implying the success of an assay when illustrated using model systems, rather than with the plant genomic target DNAs. The GM motifs selected for our studies were within Roundup ReadyTM Soya (RRS) and MON810 maize. Although knowledge of the target sequence is a prerequisite for the function of this assay, the potential of using RCA is explored.
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References
Demidov VV (2002) Expert Rev Mol Diagn 2:542–548
Pickering J, Bamford A, Godbole V, Briggs J, Scozzafava G, Roe P, Wheeler C, Ghouze F, Cuss S (2002) Nucleic Acids Res 30:e60
Alsmadi OA, Bornarth CJ, Song W, Wisniewski M, Du J, Brockman JP, Faruqi AF, Hosono S, Sun Z, Du Y, Wu X, Egholm M, Abarzua P, Lasken RS, Driscoll MD (2003) BMC Genomics 4:21–38
Larsson C, Koch J, Nygren A, Janssen G, Raap AK, Landergren U, Nilsson M (2004) Nat Methods 1:227–232
Burns MJ, Valdivia H, Harris N (2004) Anal Bioanal Chem 378:1616–1623
Taverniers I, Windels P, Van Bockstaele E, De Loose M (2001) Eur Food Res Technol 213:417–424
Hubner P, Waiblinger HU, Pietsch K, Brodmann P (2001) J AOAC Int 84:1855–1864
Hernandez M, Pla M, Esteve T, Prat S, Puigdomenech P, Ferrando A (2003) Transgenic Res 12:179–189
Taverniers I, Van Bockstaele E, De Loose M (2004) Anal Bioanal Chem 378:1198–1207
Lizardi PM, Huang X, Zhu Z, Bray-Ward P, Thomas DC, Ward DC (1998) Nat Genet 19:225–232
Nelson JR, Cai YC, Giesler TL, Farchaus JW, Sundaram ST, Ortiz-Rivera M, Hosta LP, Hewitt PL, Mamone JA, Palaniappan C, Fuller CW (2002) Biotechniques Suppl:44–47
Frieden M, Pedroso E, Kool ET (1999) Angew Chem Int Ed Engl 38:3654–3657
Dean FB, Nelson JR, Giesler TL, Lasken RS (2001) Genome Res 11:1095–1099
Westin L, Xu X, Miller C, Wang L, Edman CF, Nerenberg M (2000) Nat Biotechnol 18:199–204
Nallur G, Luo C, Fang L, Cooley S, Dave V, Lambert J, Kukanskis K, Kingsmore S, Lasken R, Schweitzer B (2001) Nucleic Acids Res 29:e118
Wang G, Maher E, Brennan C, Chin L, Leo C, Kaur M, Zhu P, Rook M, Wolfe JL, Makrigiorgos GM (2004) Genome Res 14:2357–2366
Faruqi AF, Hosono S, Driscoll MD, Dean FB, Alsmadi O, Bandaru R, Kumar G, Grimwade B, Zong Q, Sun Z, Du Y, Kingsmore S, Knott T, Lasken RS (2001) BMC Genomics 2:4
Nilsson M, Gullberg M, Dahl F, Szuhai K, Raap AK (2002) Nucleic Acids Res 30:e66
Qi X, Bakht S, Devos KM, Gale MD, Osbourn A (2001) Nucleic Acids Res 29:e116
Detter JC, Jett JM, Lucas SM, Dalin E, Arellano AR, Wang M, Nelson JR, Chapman J, Lou Y, Rokshar D, Hawkins TL, Richardson PM (2002) Genomics 80:691–698
Dahl F, Baner J, Gullberg M, Mendel-Hartvig M, Landegren U, Nilsson M (2004) Proc Natl Acad Sci U S A 101:4548–4553
Esteban JA, Salas M, Blanco L (1993) J Biol Chem 268:2719–2726
de Vega M, Blanco L, Salas M (1998) J Biol Chem 273:28966–28977
Szemes M, Bonants P, de Weerdt M, Baner J, Landegren U, Schoen CD (2005) Nucleic Acids Res 33:e70
Baner J, Isaksson A, Waldenstrom E, Jarvius J, Landegren U, Nilsson M (2003) Nucleic Acids Res 31:e103
Hardenbol P, Yu F, Belmont J, Mackenzie J, Bruckner C, Brundage T, Boudreau A, Chow S, Eberle J, Erbilgin A, Falkowski M, Fitzgerald R, Ghose S, Iartchouk O, Jain M, Karlin-Neumann G, Lu X, Miao X, Moore B, Moorhead M, Namsaraev E, Pasternak S, Prakash E, Tran K, Wang Z, Jones HB, Davis RW, Willis TD, Gibbs RA (2005) Genome Res 15:269–275
Hardenbol P, Baner J, Jain M, Nilsson M, Namsaraev EA, Karlin-Neumann GA, Fakhrai-Rad H, Ronaghi M, Willis TD, Landegren U, Davis RW (2003) Nat Biotechnol 21:673–678
Acknowledgements
This work was supported by the United Kingdom Food Standards Agency. We thank Mr. Hernan Valdivia for extracting genomic DNA from soybean and maize-certified reference materials.
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Pang, S., Qureshi, F., Shanahan, D. et al. Investigation of the use of rolling circle amplification for the detection of GM food. Eur Food Res Technol 225, 59–66 (2007). https://doi.org/10.1007/s00217-006-0382-1
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DOI: https://doi.org/10.1007/s00217-006-0382-1