TY - JOUR
T1 - Leaf-punch method to prepare a large number of PCR templates from plants for SNP analysis
AU - Shiokai, Sachiko
AU - Kitashiba, Hiroyasu
AU - Shirasawa, Kenta
AU - Nagano, Kuniaki
AU - Nishio, Takeshi
N1 - Funding Information:
Acknowledgments This work was supported in part by a Grant-in-Aid for Scientific Research (A) (no. 19208001) from the Japan Society for the promotion of science (JSPS). CSSL228 was provided by Rice Genome Resource Center, Japan. S. Shiokai is a recipient of a research fellowship from the Japan Society for the promotion of science for young Scientists.
PY - 2009/2
Y1 - 2009/2
N2 - DNA preparation is indispensable for genotyping by DNA polymorphism analysis, and that for a large number of plants is laborious. In the present study, a small leaf disk of rice, 1-2 mm in diameter, punched by a mini cork borer was found to be directly usable as a PCR template. DNA fragments <300 bp were amplified efficiently. Leaf disks of 1-1.5 mm in diameter were better than those of 2 mm for a small volume of reaction mixture. Multiplex PCR was possible with four or eight primer pairs using the small leaf disk as a template. Leaf disks of Arabidopsis, Lotus, wheat, soybean, tomato, Chinese cabbage, and melon were also good PCR templates. This method for preparation of PCR templates, named the leaf-punch method, was applicable to SNP analysis of a large number of plants by dot-blot-SNP analysis.
AB - DNA preparation is indispensable for genotyping by DNA polymorphism analysis, and that for a large number of plants is laborious. In the present study, a small leaf disk of rice, 1-2 mm in diameter, punched by a mini cork borer was found to be directly usable as a PCR template. DNA fragments <300 bp were amplified efficiently. Leaf disks of 1-1.5 mm in diameter were better than those of 2 mm for a small volume of reaction mixture. Multiplex PCR was possible with four or eight primer pairs using the small leaf disk as a template. Leaf disks of Arabidopsis, Lotus, wheat, soybean, tomato, Chinese cabbage, and melon were also good PCR templates. This method for preparation of PCR templates, named the leaf-punch method, was applicable to SNP analysis of a large number of plants by dot-blot-SNP analysis.
KW - DNA preparation
KW - Large-scale genotyping
KW - Marker-assisted selection
KW - PCR template
KW - SNP analysis
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U2 - 10.1007/s11032-008-9244-9
DO - 10.1007/s11032-008-9244-9
M3 - Article
AN - SCOPUS:58149189828
SN - 1380-3743
VL - 23
SP - 329
EP - 336
JO - Molecular Breeding
JF - Molecular Breeding
IS - 2
ER -