Method for rapidly identifying H. villosa chromosomes in wheat background
A tufted wheat and chromosome technology, applied in the field of molecular cytogenetics, can solve the problems of inability to identify rye chromosomes, complicated processes, and inability to be clear at a glance, and achieves the effects of overcoming cumbersome preparation, low cost and easy operation.
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Embodiment 1
[0048] Example 1 Analysis of Wheat-Viligal repeats and Determination of Oligonucleotide Probe Sequences
[0049] According to the characteristics of the distribution of transposable repeat sequences in the plant genome in a scatter manner, we start with the analysis of transposable repeat sequences and search for oligonucleotide probes. Based on the fact that there are relatively few transposable repetitive sequences isolated and cloned from the genome of T. villosa, and based on the principle that each species of wheat crops has a certain genetic relationship, consider using the method of sequence transfer between species to develop suitable oligonucleotides acid probe.
[0050] 1. Determination of oligonucleotide probe nucleotide sequence and probe design
[0051] (1) Collect all the reported transposable repeat sequences isolated and cloned from the rye genome and the reported rye-specific oligonucleotide probe sequences.
[0052] From the NCBI database, download all publ...
Embodiment 2
[0060] Example 2 Establishment of the Chromosomal Identification Method for T. villosa in the Wheat Background
[0061] Two kinds of oligonucleotide probes (Oligo-Ku and (GT) determined in embodiment 1 7 ) were mixed, and ND-FISH was used to identify the chromosomes of A. villosa in the wheat background. The method described in (Han et al.2006) was used for the preparation of the root tip metaphase chromosomes of wheat-T. villosa amphidiploid and wheat cultivar Mianmai 37, and the method described in (Fu et al.2015) for the ND-FISH analysis process . details as follows:
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