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Bulletin of Botanical Research ›› 2000, Vol. 20 ›› Issue (2): 195-200.

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MICRODISSECTION AND AMPLIFICATION IN VITRO OF CHROMOSOME 1R IN RYE (Secale cereale L.) AND DETERMINATION OF THE AMPLIFIED PRODUCTS

WEI Ji-cheng, SHI Rui, GUO Chang-hong, GUO Dong-lin, MA Xu-jun, WANG Tong-chang   

  1. Harbin Normal University, Harbin 150040
  • Received:1998-09-21 Online:2000-06-15 Published:2016-06-14

Abstract: The chromosome 1R of rye was microdissected and isolated under reverse-microscope with a manipulator from the cells of root tip at mitotic metaphase. Single chromosome 1R was then transferred into 0.5 ml centrifuge tube. After the treatment with Protease K, the DNA of chromosome 1R was digested with restriction enzyme Sau3A, then the Sau3A linker-adaptor was ligated to the ends of the DNA fragments, which was subsequently amplified by means of linker-adapter polym erase chain reaction (LA-PCR) with one of chains of the linker-adapter as primer. By dot blotting with biotinlatyled rDNA of wheat and genome DNA of rye as probes, it was confirmed that the amplified products by LA-PCR are derived from the DNA of chromosome 1R in rye. This work would facilitated to construct the sub-genomic DNA library of chromosome 1R and to screen the specific probes of chromosome 1R in rye.

Key words: rye, chromosome 1R, microdissection, LA-PCR, rDNA