Recombined nucleic acid fragment reccr010007 and its detection primers and applications
A technology of fragments and recombinant single plants, applied in the field of molecular biology, can solve the problems of time-consuming and low efficiency
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Embodiment 1
[0075] Example 1 Breasting recombinant plants introduced into the rice blast genome fragment
[0076] The materials used in this embodiment are rice 'deep 95b' and rice 'China 3418b'.
[0077] Rice 'China 3418B' has a good rice blast resistance, and it is speculated that the region of PI1 and PIK cluster alleles in chromosome, which may play a key role in the region of the rice blast resistance of the material.
[0078] During the breeding process of recombinant plants, the reassembly selection was performed using molecular markers, and the foreground selection molecular marker used was screened. Referring to Rice Japan Chinenome MSU / TIGR Note No. 6.1, the 11th chromosome 27, 155,000 to 28, 495,000 DNA sequences were downloaded. The SSR bit point in the above sequence is scanned using the SSRLocator software. Using Primer Premier 3.0 software to design primers for SSR sites to find primers 385. Through the method of PCR, the above primers are screened in the polymorphism of 'Chin...
Embodiment 2
[0088] Example 2 Improve the identification of homologous recombinant fragments after rice blast resistance genomic segment
[0089] In order to determine the size of the imported rice blast resistance genomic fragment, the homozygous single strain of the "deep 95b 'introduction fragment was sequenced. Sequencing of the homologous recombination fragment of the target genomic fragment. The anti-rice disease generated recombinant nucleic acid fragment contained in CR010007 was named RECCR010007.
[0090] The RICE60K test results were initially determined by rice total genome breeding chip, and RECCR010007 is located upstream of SNP marking F1127982620TC.
[0091] At the same time, three samples of 'deep 95b', 'China 3418b' and Cr010007 were used to use Miseq sequencing technology. The library was established using the Truseq Nano DNA LT Kit (Illumina) kit, using the libraryquantification kit-universal (kapa biosystems) kit, using the MiseQv2Reagent Kit (Illumina) kit for sequencing ...
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