Insertion of a reamplification round into the ISSR-PCR protocol gives new flax fingerprinting patterns

• Autorzy: Wiesner I , Wiesnerova D.
• Języki publikacji: EN

Abstrakty

EN

We expanded the basic ISSR-PCR protocol by an additional PCR reamplification round in order to detect whether increased PCR productivity would give new bands in ISSR patterns. We found that the reamplification step had a prominent impact on the quality of the inter-simple-sequence repeat (ISSR) PCR patterns of flax, depending on the particular primer used for PCR amplification. We could clearly distinguish between two types of reamplification effect. Most ISSR primers (16 out of 21) gave no reamplification effect as usual, but five primers (23.8%) provided a new ISSR fingerprinting pattern after the 2nd reamplification round, leaving the previous 1st round pattern completely blank. Therefore, we recommend the expansion of a basic ISSR-PCR protocol for another reamplification round in order to mine out full the fingerprinting potential from ISSR-PCR method.

Słowa kluczowe

EN

germplasm; fingerprinting; Linum usitatissimum; flax; polymerase chain reaction; genetic marker; identification

• Wydawca: -
• Czasopismo: Cellular and Molecular Biology Letters
• Rocznik: 2003
• Tom: 08
• Numer: 3
• Opis fizyczny: p.743-748,fig.

Twórcy

autor

Wiesner I

• Academy of Sciences of the Czech Republic, Branisovska 31, Ceske Budejovice, CZ 370 05, Czech Republic

autor

Wiesnerova D.

Bibliografia

• 1.Zietkiewicz, E., Rafalski, A. and Labuda, D. Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics 20 (1994) 176-183.
• 2.Wiesner, I., Wiesnerová, D. and Tejklová, E. Effect of anchor and core sequence in microsatellite primers on flax fingerprinting patterns. J. Agricult. Sci. 137 (2001) 37-44.
• 3.Hengen, P.N. Reamplification of PCR fragments. Trends Biochem. Sci. 20 (1995) 124-125.
• 4.Arnau, J., Housego, A.P. and Oliver, R.P. The use of RAPD markers in the genetic analysis of the plant-pathogenic fungus Cladosporium fulvum. Cur. Genet. 25 (1994) 438-444.
• 5.Nath, K., Sarosy, J.W., Hahn, J. and Di Como, Ch. J. Effects of ethidium bromide and SYBR Green I on different polymerase chain reaction systems. J. Biochem. Biophys. Meth. 42 (2000) 15-29.
• 6.Pierre, C., Lecossier, D., Boussougant, Y., Bocart, D., Joly, V., Yeni, P. and Hance, A.J. Use of a reamplification protocol improves sensitivity of detection of Mycobacterium tuberculosis in clinical samples by amplification of DNA. J. Clin. Microbiol. 29 (1991) 712-717.
• 7.Chaiprasert, A., Prammanan, T., Samerpitak, K., Pattanakitsakul, S., Yenjitsomanus, P., Jearanaisilawong, J. and Charoenratanakul, S. Direct detection of M. tuberculosis in sputum by reamplification with 16S rRNA- based primer. Asia-Pacif. J. Mol. Biol. Biotech. 4 (1996) 250-259.
• 8.Wang, H., Xia, Z.B., Wang, Y.H., Li, C.H. and Sun, L.Y. Recovering and reamplifying of the differentially expressed cDNA bands isolated from mRNA differential display - a modified method. Mol. Biotechnol. 9 (1998) 171-173.
• 9.Godwin, I.D., Aitken, E.A. and Smith, L.W. Application of inter simple sequence repeat (ISSR) markers to plant genetics. Electrophoresis 18 (1997) 1524-1528.