High-throughput tissue grinder for analysis of purity of pumpkin hybrids

In rice, corn, soybean, rape and other important crops, SSR marking technology has become a commonly used purity identification method. Because of its abundant number, high polymorphism, codominant inheritance, simple operation and stable and reliable results, SSR markers have become the ideal marker type for the purity identification of crop varieties. Rapid and simple SSR detection methods have also been researched and established. "The development of pumpkin SSR markers is late, and it is mainly used for the construction of genetic linkage maps, gene mapping and other basic research, and there are few applied researches. High-throughput tissue grinding The instrument can play an important role in researching sample types.

After soaking the seeds for 1 h, place them in a germinating box with filter paper, 37e, and pay attention to moisturizing. About 5 to 6 days of germination, take the embryonic axis of a single-grained seed about .05 cm, and place it in a 20 mL centrifuge tube. Grinding with a tissue grinder; adding 600 liter LCATB extract, 65e water bath 10mni; adding equal volume of chloroform: isoamyl alcohol (24:1), fully inverting and mixing for about 10 min, centrifuging at 120001 丫 11 ]in for 10 min; The serum was mixed with an equal volume of pre-cooled isopropanol (inverted gently), left at room temperature for about 30 minutes, centrifuged at 1200 Ori for 10 min, and the supernatant was discarded and inverted to allow it to air dry or be placed in a clean atmosphere. Let it dry on the workbench and add appropriate amount of ddHZO to dissolve it."

Through the participation of high-throughput tissue grinders, the simple SSR detection system is simpler and more practical than the conventional SSR detection system, but there are also a small amount of extracted genomic DNA! The disadvantage of not being easy to store for a long time. "Also, although this study uses The detection system greatly reduces the amount of reagents compared to the conventional detection system. However, in theory, the PCR reaction system can be further reduced under the premise of guaranteeing the sample quantity; in order to improve the detection efficiency, multiple PCRs can also be explored so as to simultaneously The difference in multiple sites is detected. Therefore, the amplification system can be further optimized. "At the time of electrophoresis detection, this study was only spotted once. For the amplification products of suitable primers, two or more spots can be explored to facilitate Saving the cost of making glue, the work in this area still needs further exploration based on the size of the amplified product.

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