DESIGN OF ELECTROPHORESIS DEVICE FOR OPTIMATION OF DNA VISUALIZATION AND DNA CONCENTRATION USING SOFTWARE
(1) Jl. Prof. Soedarto, UNDIP, Tembalang, Semarang. 50275
(2) Physics Laboratory, Faculty of Mathematics and Natural Sciences, Diponegoro University, Semarang, Indonesia
(3) Physics Laboratory, Faculty of Mathematics and Natural Sciences, Diponegoro University, Semarang, Indonesia
(4) Physics Laboratory, Faculty of Mathematics and Natural Sciences, Diponegoro University, Semarang, Indonesia
Abstract
Molekul DNA menunjukkan polarisasi yang kuat sehingga memungkinkan baik gerak elektroforesis berdasarkan muatan negatifnya maupun gerak dielektroforesis berdasarkan induksi polarisasi. Perancangan alat menggunakan kombinasi prinsip elektroforesis dan dielektroforesis dilengkapi perangkat lunak untuk mengukur konsentrasinya sangat diperlukan. Utamanya mengingat uji kualitatif DNA berbasis visualisasi pada gel elektroforesis bersifat sangat subyektif dan kurang terukur. Pengukuran konsentrasi DNA menggunakan spektrofotometer UV/VIS sangat tergantung oleh ketersediaannya di laboratorium. Penelitian bertujuan untuk mendesain piranti untuk mengukur konsentrasi DNA berdasarkan visualisasinya pada gel elektroforesis menggunakan perangkat lunak berbasis MatLab. Pengukuran konsentrasi DNA didasarkan visualisasinya pada gel elektroforesis lalu dibandingkan dengan hasil penghitungan spektrofotometer UV/VIS. Hasil penelitian menggunakan piranti tersebut memperlihatkan visualisasi DNA yang lebih optimal. Hasil pengukuran jumlah DNA menggunakan spektrofotometer memiliki kecenderungan yang sama dengan hasil pengukuran menggunakan perangkat lunak berbasis MatLab meskipun terdapat perbedaan nilai kuantitatif.
ABSTRACT
Molecules of deoxyribo nucleic acid (DNA) show a strong polarization allowing for both motions of the dielectrophoresis induced by polarization and electrophoresis based on its negative charge. Considering high subjective and less quantifiable result of the visualization based qualitative test of DNA on gel electrophoresis, designing the tool using a combination of the principles of electrophoresis and dielectrophoresis completed with a software for optimization of DNA visualization and to measure the concentration of small and large–sized DNA fragment is very needed. Accuracy of measurement of DNA concentration using a spectrophotometer UV /VIS is depend on its availability in the laboratory. The aim of this study was to design device for optimization of DNA visualization and measuring the concentration in the gel electrophoresis using MatLab- based software. Experiment using this software measured the concentration of DNA based on its visualization and compared it with calculation obtained from spectrophotometer UV/VIS. The research results showed that the amount of DNA analysed using a spectrophotometer tend to similar with the measurement results using the MatLab-based software although there was differences in quantitative values.
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Alberts B, D.Bray, J Lewis , M. Ralf, K. Roberts, and JD Watson. (1991). Molecular Biology of The Cell. Garland Publ. London.
Ausubel, F., R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith, K. Struhl. (1995). Short Protocols in Molecular Biology. A Compedium of Methods from Current Protocols in Molecular Biology. 3nd Ed . Wiley & Sons. Inc. USA. 2-10.
Bakewell D.J. and H. Morgan. (2006). Dielectrophoresis of DNA: time- and frequency-dependent collections on microelectrodes. IEEE Trans Nanobioscience 5(2) : 139 – 146
Castellarnau M., Errachid A., Madrid C., Juárez A.and J. Samitier. (2006).
Dielectrophoresis as a Tool to Characterize and Differentiate Isogenic Mutants of Escherichia coli. Biophysical Jour., 91(10) : 3937-3945
Chou, C, Tegenfeldt J.O., Bakajin O., Chan S.S.,. Cox E.C, Darnton N., Duke T. and R.H. Austin. (2002). Electrodeless Dielectrophoresis of Single- and Double-Stranded DNA. Abstract. Biophysical Jour. 83 (4) : 2170-2179
Day, R. A. dan A. L. Underwood. (2002). Analisa Kimia Kuantitatif. Erlangga, Jakarta.
Gemen S. and D. Gemen. (1984). Stochastic Relaxation, Gibbs distributions and Bayesian Restoration. IEEE Transactions of Pattern Analysis and Machine Intelligence, 6 721
Goldfarb D and Yin W (2005). Second-Order Cone Programing Methods For Total Variation Based Image Restoration. AMS.
Haario, H., Laine, M., Lehtinen, M., Saksman, E. and Tamminen, J. (2004). Markov chain Monte Carlo methods for high dimensional inversion in remote sensing. J. R. Statist. Soc. B 66(3), 591-608
Hoeb M., Rädler J.O., Klein S., Stutzmann M. and M.S. Brandt. (2007). Light-Induced Dielectrophoretic Manipulation of DNA. Biophysical Jour., 93(3): 1032-1038
Kua, C.H, Lam Y.C., Yang , C. and K. Youcef-Toumi. (2004). Review of bio-particle manipulation using dielectrophoresis. Nanyang Tech Univ, Singapore.; Massachusetts Inst. of Tech., Cambridge, Massachusetts.p. 1-7
Lapizco-Encinas, B.H., Simmons, B.A, Cummings, E.B. and Y. Fintschenko. (2004). Insulator-based dielectrophoresis for the selective concentration and separation of live bacteria in water. Electrophoresis 25 : 1695–1704
Miles R., Wang A., Bettencourt K., Nasarabadi S., Belgrader P., Davidson J.C. and R.P. Mariella Jr. (1989). Manipulation of DNA for Use in Microfluidic Devices. UCRL-JC-132459 Preprint. Lawrence Livermore. Nat. Lab.
Murthy K. P. N. (2005). Bayesian Restoration of Digital Images Employing Markov Chain Monte Carlo”, eprint arXiv:cs/0504037
Regtmeier J., Duong T.T., Eichhorn R., Anselmetti R, and A. Ros. (2007). Dielectrophoretic Manipulation of DNA: Separation and Polarizability. Anal. Chem., 79(10) : 3925 -3932
Regtmeier J., Eichhorn R., Bogunovic L., A. Ros and Anselmetti D. (2010). Dielectrophoretic Trapping and Polarizability of DNA : The Roll of Spatial Conformation. Anal. Chem., 82:7141-7149
Sambrook, J., Fritsch, E. R. & T. Maniatis.(1989). Molecular Cloning A Laboratory Manual. Second Edition. Cold Spring Harbor Laboratory Press, USA.
Tanaka K and N Yoshiike. (2003). Statistical-Mechanical Approach to Probabilistic Image Processing. Workshop on Statistical Mechanical Approach to Probabilistic Information Processing.
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