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Message - 7th IAL Symposium

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The 7 th International Association for Lichenology <strong>Symposium</strong> 2012<br />

(3A-P24) Submission ID: <strong>IAL</strong>0299-00002<br />

SIMPLE TECHNIQUE FOR ON SITE IDENTIFICATION OF PYXINE COCOES (Sw.) NYL.,<br />

A FOLIOSE LICHEN, BASED ON GENOSENSOR<br />

Kosakul T. 1 , Chaumpluk P. 1 , Artchawakom T. 2<br />

1 Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok, Thailand<br />

2 Sakaerat Environmental Research Station, Thailand Institute of Scientific and Technological Research,<br />

Nakornratchasima Province, Thailand<br />

A simple technique for an on-site identification of Pyxine cocoes (Sw.) Nyl., a folliose lichen, based<br />

on variable sequence domains within the internal transcribed spacer (ITS) region had been developed. This<br />

technique involved an isothermal DNA amplification of 84 nucleotides fragment of target ITS element for DNA<br />

signals multiplication at 63 ºC for 2 hours and a signal detection via visual fluorescence signals simultaneously<br />

using DNA binder. Results of the detection can be observed with unaided eye within 2 hours without the need of<br />

thermocycler and gel electrophoresis. DNA signals were detected only from Pyxine cocoes but not from Chrysothrix<br />

xanthina Kalb. and Parmotrema tinctorum (Nyl.) Hale. The detection limit was at 50 copies of target DNA<br />

domains per reaction, equally to 25 cells. This constitutes a basis for a simple and accurate detection of target<br />

lichen species, suitable for field application.<br />

165<br />

3A-P

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