A method for identifying the genetic purity of pear hybrid seedlings
An identification method and seedling technology, applied in the biological field, can solve problems such as time-consuming, low accuracy, and difficulty in distinguishing, and achieve the effects of improving identification efficiency, low cost, and stable labeling
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Embodiment 1
[0030] In the present invention, the female parent 'Hua Su', the male parent 'Cui Guan' and 'Su Cui No. 1' are all provided by Jiangsu Academy of Agricultural Sciences. Example 1: Rapid identification of genetic purity of 'Su Cui No. 1' hybrid seedlings.
[0031] The rapid identification of the genetic purity of pear 'Sucui No. 1' hybrid seedlings using SRAP molecular marker technology, the main operating procedures are as follows:
[0032] 1. DNA extraction:
[0033] (1) Take about 0.1 g of young leaves of pear 'Sucui No. 1', grind with liquid nitrogen, add 500 μl of CTAB lysate (2% CTAB, 2mol / L NaCl, 20mmol / L EDTA, 100mmol / L Tris-HCl (pH =8.0) and 0.2% β-mercaptoethanol (volume ratio) into a 1.5ml centrifuge tube, in a 65°C water bath for 30-50min;
[0034] (2) Take out and centrifuge at 16°C, 12,000rpm for 10min, transfer the supernatant into a freshly sterilized 1.5ml centrifuge tube, add an equal volume of chloroform:isoamyl alcohol (24:1, volume ratio) mixture, Shake ...
Embodiment 2
[0050] The method for extracting the genome used in this example, the PCR method and the detection method are the same as Example 1, the difference is that this example uses the co-dominant SRAP primer combination JAASSRMe3 / JAASSREm11 to pear seedlings of 10 other varieties for identification ( figure 2 ), PCR results ( image 3 ) shows that the pear seedlings of other varieties have obvious polymorphic amplification bands, but compared with the hybrid seedlings of 'Sucui 1', only 'Sucui 1' has specific bands at 60bp and 90bp 60bp maternal-specific marker band and 90bp paternal-specific marker band ( image 3 ), therefore, the results of this experiment show that the SRAP molecular marker method has strong specificity and reliability, and can be used to identify the genetic purity of pear hybrid seedlings.
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