Expression detection method for sweet sorghum sucrose phosphate synthase gene and amplification primer
A technology for phosphate synthase and gene amplification, which is applied in the field of amplification primers, sweet sorghum sucrose phosphate synthase gene, and expression detection of sweet sorghum sucrose phosphate synthase gene, which can solve the problems of no comprehensive reports and achieve band The effects of clearness, shortening the breeding cycle, and overcoming difficulties in phenotypic identification
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[0075] Example 1: Correlation analysis of sweet sorghum sucrose phosphate synthase (SPS) activity and sucrose content
[0076] The present invention measures the F of male parent, female parent and their hybrids at different periods. 5 The activity of SPS in leaves of high-sugar population and low-sugar population, to understand the changing law of SPS between different populations in different periods. Parents and F 5 The activity of SPS in leaves of high-sugar population and low-sugar population is gradually increasing. At the maturity stage, the activities of the three sweet sorghum varieties, the female parent, the high-sugar population and the low-sugar population, reached their maximum values of 14.86, 17.63, and 13.76 μmol / hgFW respectively; the male parent’s SPS activity at the mature stage was slightly higher than at the heading stage The drop. There is not much difference in the SPS activity of the four varieties at the seedling stage, but there is a gap in the SPS a...
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[0077] Implementation Example 2: Analysis of the extraction results of sweet sorghum RNA
[0078] Agarose gel electrophoresis detection showed that the RNA extracted by the modified Trizol method had 3 bands. The 28S band in the total RNA was about twice as bright as the 18S band, indicating that it was less degraded during the extraction process. The integrity of the extracted RNA is very good. The 5S band is relatively shallow, indicating that there is no contamination of RNA and can be used as a template in the next experiment.
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[0079] Implementation Example 3: Primer Design of Sucrose Phosphate Synthase (SPS)
[0080] The primers in Table 1 are designed based on a known cDNA sequence of the coding region of the sweet sorghum SPS3-1 gene, using Oligo6 software and Primer primier software. Named SPS-1, β-Actin is the housekeeping gene, and the lengths of the target fragments amplified by the two pairs of primers are 121bp and 210bp, respectively.
[0081] Table 1 Primers used for SPS gene amplification
[0082]
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