Leaf-punch method to prepare a large number of PCR templates from plants for SNP analysis

被引：7

作者：

Shiokai, Sachiko ^{[1
]}

Kitashiba, Hiroyasu ^{[1
]}

Shirasawa, Kenta ^{[1
]}

Nagano, Kuniaki ^{[2
]}

Nishio, Takeshi ^{[1
]}

机构：

[1] Tohoku Univ, Grad Sch Agr Sci, Aoba Ku, Sendai, Miyagi 9818555, Japan

[2] Miyagi Prefectural Furukawa Agr Expt Stn, Furukawa, Miyagi 9896227, Japan

来源：

MOLECULAR BREEDING | 2009年 / 23卷 / 02期

基金：

日本学术振兴会;

关键词：

DNA preparation; PCR template; SNP analysis; Large-scale genotyping; Marker-assisted selection; SINGLE-NUCLEOTIDE POLYMORPHISMS; RICE ORYZA-SATIVA; GENE-EXPRESSION; IDENTIFICATION; DISPERSAL; RUFIPOGON; TRAITS; POLLEN; VIRUS; LINES;

D O I：

10.1007/s11032-008-9244-9

中图分类号：

S3 [农学（农艺学）];

学科分类号：

0901 ;

摘要：

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.

引用

页码：329 / 336

页数：8

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