Construction method of livestock ingestion composition correction model based on macro bar code
A technology for constructing methods and calibrating models, which is applied in the field of molecular biology, can solve problems such as deviations in measurement results, and achieve the effect of improving accuracy
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Embodiment 1
[0034] Embodiment 1 is to the mensuration of pasture component in simulated ration
[0035] After the collected 9 kinds of pastures were fully ground with liquid nitrogen, they were fully mixed according to the set ratio (dry matter ratio) and used as a simulated diet. The ratio is shown in Table 1 below. Including 10 treatments, each treatment repeated 4 times, a total of 40 forage mixture samples. About 0.5g of each mixed forage sample was loaded into a centrifuge tube and stored in a liquid nitrogen tank.
[0036] Table 1 Composition of simulated diet
[0037]
[0038] The simulated diet samples were ground and mixed with liquid nitrogen, and the total DNA of the samples was extracted with a plant genomic DNA extraction kit (DNAsecurePlant Kit). Using the above DNA as a template, use primers rD5-ITS2 and rb1-ITS2f (Table 2) reported in the literature to PCR amplify the plant ITS2 sequence, and add an 8bp tag sequence to each sample for distinguishing sequencing analysi...
Embodiment 2
[0048] Example 2 Regression Model Construction and Accuracy Inspection of Simulated Dietary Components
[0049] Linear regression analysis was used to determine the quantitative relationship between the relative proportions of sequenced DNA sequences and the forage proportion in the simulated diet, and the resulting regression equation was used to correct direct estimates.
[0050] Regression correction was performed on the direct estimation results of feeding components obtained from high-throughput sequencing, and the accuracy of the estimated and corrected results was evaluated by the Kulczyński similarity coefficient (KSI, %). Its calculation formula is as follows:
[0051]
[0052] Among them, c i is the smaller value between the real value and the estimated value of the ith forage in the simulated diet; (a i +b i ) is the sum of the ratios of true and estimated values for each forage component.
[0053] The sequence number of each OTU was obtained after Illumina...
Embodiment 3
[0059] Example 3 Estimation and Correction of Diet Components for Housed Sheep
[0060] Using the method of ITS2 barcode sequencing in the present invention, the feed intake components of sheep were estimated. The accuracy of the method was also tested using KSI ( Image 6 ). The results of variance analysis showed that dietary treatment, regression equation correction method and the interaction between them had a very significant effect on KSI (P<0.001). After correction by the two regression equations, KSI increased significantly (P<0.001). Comparing the three estimated results, it can be found that the KSI is the highest (88.41%) after the second correction equation (CE-2), followed by the second correction equation CE-1 (70.24%), without The corrected KSI value was the lowest (57.97%).
[0061] From the perspective of dietary components, the similarity between the estimated value and the real value was low (46.98%-67.99%), and the similarity between the estimated value...
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