NOTEBOOK
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Date |
2025.7 |
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Participants |
Wu Wenran, Du Xiaolu, Kang Qinhao, Chen Ziqi, Zhang Fengshang, Pan Yanxi, Li Xinyang, Wang Youjie, Wang Zibing, Che Kaiyue, Zhou Yuanrui |
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Contents |
1.Prepare LB solid and liquid media, followed by high-temperature sterilization. 2.Perform PCR amplification of the target genes PDI and Thaumatin-A/B/C/D, and retain the amplified products for subsequent use after a 2-hour amplification reaction. 3.Prepare agarose gel and set it aside after solidification. 4.Inoculate the pRSFDuet plasmid into competent cells for shake culture. |
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Results |
Agarose gel electrophoresis results of PCR for five clones |
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Date |
2025.7 |
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Participants |
Wu Wenran, Du Xiaolu, Kang Qinhao, Chen Ziqi, Zhang Fengshang, Pan Yanxi, Li Xinyang, Wang, Youjie, Wang Zibing, Che Kaiyue, Zhou Yuanrui, Wu Xinke |
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Contents |
1.Extraction of pRSFDuet Vector Plasmid 2.Restriction Enzyme Double Digestion 3.DNA Gel Electrophoresis Verification 4.Gel Extraction of Linearized Vector DNA 5.Homologous Recombination pRSFDuet-PDI 6.Heat-Shock Transformation E.coil DH5α 7.Make kan+ Antibiotic-Resistant Plate 8.DNA Gel Dissolution and Purification |
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Results |
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Date |
2025.7 |
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Participants |
Du Xiaolu, Wu Wenran, Kang Qinhao, Chen Ziqi, Pan Yanxi, Wang Youjie, Wu Xinke, Zhou Yuanrui |
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Contents |
1.prepare LB liquid/solid (solid:4×100ml) (liquid:4×150ml) 2.Colony PCR identification (picking 8 groups of colony, PCR identification 3.Identify the correct group of colony and prepare for expanding cultivation 4.Agarose gel electrophoresis determine the molecular weight of nucleic acid 5.shaking culture let the pRSF-PDI to grow |
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Results |
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Date |
2025.7 |
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Participants |
Wu Wenran, Chen Ziqi, Wu Xinke, Zhou Yuanrui, XU Yihan, Huang Yunni, Che Kaiyue |
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Contents |
1.Extraction of PRSF-PDI Vector Plasmid 2.Restriction Enzyme Double Digestion 3.Prepare agarose gel and set it aside after solidification. 4.alidation of PRSF-PDI linear by agarose gel electrophoresis 5.Agarose gel recovery 6.Homologous Recombination 7.Heat-Shock Transformation |
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Results |
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Date |
2025.8 |
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Participants |
Du XiaoLu, Chen Ziqi, Wu Xinke, Zhou YuanRui, XU Yihan, Huang Yunni, Che Kaiyue |
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Contents |
1.pRSFDuet-Thaumatin-A/B/C/D colony PCR identification 2.Prepare agarose gel and set it aside after solidification. (15%PAGE Gel) 3. Enlarging cultvate of E. coli DH5a and Origami2(DE3) |
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Results |
Agarose gel electrophoresis results of Colony PCR identification for five clones
Agar plate showing BL21 colonies after transformation |
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Date |
2025.8 |
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Participants |
Du XiaoLu, Chen Ziqi, Wu Xinke, Zhou YuanRui, XU Yihan, Huang Yunni, Che Kaiyue, Kang Qinhao, Zhang Fengshang |
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Contents |
1.Plasmid extraction and identification 2.Bacterial culture 3.Agarose gel and electrphoresis |
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Results |
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Date |
2025.8 |
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Participants |
Du Xiaolu, Kang Qinhao, Zhang Fengshang, Pan Yanxi, Li Xinyang, Wang Zibing, Che Kaiyue, Zhou Yuanrui, Pan Yanxi, Wu Xinke |
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Contents |
1.Extract 4 kinds of crude proteins by ultrasonic disruption 2.Extract pure proteins by nickel column affinity purification 3.Perform SDS-PAGE gel electrophoresis to detect whether the protein size meets expectations 4.Conduct Coomassie brilliant blue decolorization and staining, and leave it overnight |
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Results |
SDS-PAGE analysis of crude protein
SDS-PAGE analysis After Protein Elution |
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Date |
2025.8 |
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Participants |
Du Xiaolu, Kang Qinhao, Chen Ziqi, Wu Xinke, Zhang Fengshang, Pan Yanxi, Wang Youjie, Che Kaiyue, Zhou Yuanrui, Huang Yunni, Xu Yihan |
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Contents |
1.Enzyme assay 2.Principles of DNA testing |
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Results |
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