Method for detecting Fenghua21 watermelon seed authenticity and purity
An authentic and rich technology, applied in the field of molecular biology, can solve the problems of indistinguishable watermelon hybrids, easy to be affected by the environment, labor-intensive and land-consuming, and achieve the effect of abundant quantity, high polymorphism and large quantity.
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Embodiment 1
[0058] Embodiment one: a kind of method for detecting the authenticity and purity of 'Fenghua 21' watermelon seeds provided by the invention comprises the following steps:
[0059] (1) Design SSR primers according to the published watermelon genome sequence information, and screen out the polymorphic and stable SSR primers in the parents and hybrids of ‘Fenghua 21’ among the designed SSR primers;
[0060] (2) Use the SSR primers finally screened in step (1) to prepare the SSR fingerprint of 'Fenghua 21';
[0061] (3) The watermelon seeds to be detected are sown, the genomic DNA of the cotyledon stage leaves is extracted by CTAB method, PCR amplification and polyacrylamide gel electrophoresis are carried out using the SSR primers finally screened out in step (1), and the DNA of the seeds to be detected is obtained. The SSR amplified band pattern is compared with the SSR fingerprint of 'Fenghua 21' prepared in step (2). If the comparison results are consistent, it can be determi...
Embodiment 2
[0067] Embodiment two: a kind of method for detecting the authenticity and purity of 'Fenghua 21' watermelon seeds provided by the invention comprises the following steps:
[0068] (1) Design SSR primers according to the published watermelon genome sequence information, and screen out the polymorphic and stable SSR primers in the parents and hybrids of ‘Fenghua 21’ among the designed SSR primers;
[0069] (2) Use the SSR primers finally screened in step (1) to prepare the SSR fingerprint of 'Fenghua 21';
[0070] (3) The watermelon seeds to be detected are sown, the genomic DNA of the cotyledon stage leaves is extracted by CTAB method, PCR amplification and polyacrylamide gel electrophoresis are carried out using the SSR primers finally screened out in step (1), and the DNA of the seeds to be detected is obtained. The SSR amplified band pattern is compared with the SSR fingerprint of 'Fenghua 21' prepared in step (2). If the comparison results are consistent, it can be determi...
Embodiment 3
[0100] Embodiment three: a kind of method for detecting the authenticity and purity of 'Fenghua 21' watermelon seeds provided by the invention comprises the following steps:
[0101] (1) Design SSR primers according to the published watermelon genome sequence information, and screen out the polymorphic and stable SSR primers in the father, mother and hybrid of 'Fenghua 21' among the designed SSR primers;
[0102] (2) Use the SSR primers finally screened in step (1) to prepare the SSR fingerprint of 'Fenghua 21';
[0103] (3) The watermelon seeds to be detected are sown, the genomic DNA of the cotyledon stage leaves is extracted by CTAB method, PCR amplification and polyacrylamide gel electrophoresis are carried out using the SSR primers finally screened out in step (1), and the DNA of the seeds to be detected is obtained. The SSR amplified band pattern is compared with the 'Fenghua 21' SSR fingerprint spectrum prepared in step (2). If the comparison results are consistent, it ...
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