Team:Nanjing high school/Notebook
Notebook
Day1
Online training to introduce the background of the subject and related knowledge.
The coach leader introduces himself and introduces the training content；
ice breaker games；
Candidates for captain election, determine the team name, logo, slogan, uniform and related surroundings；
Day2
Laboratory safety and basic operation training, molecular biology, protein and cell biology and other biological knowledge training;
Understand the common instruments and equipment in biochemical laboratories, such as centrifuges, PCR machines, sterilization pots, constant temperature shakers, etc., and practice the use of common instruments, such as micropipettes, etc.
Preparation of both liquid and sodium LB culture media, sterilized.
Cultured the strains containing pUC57-nahR plasmids.
Day3
pUC57-nahR plasmids extraction.
Preparation of Ampicillin and Kanamycin solution.
Day4
Cultured the strains containing pTrc99k plasmids (Kan), pMW119 plasmids (Amp) and Tatumella citrea.
Learn the preparation of the PCR reaction system and the use of the PCR machine, learn the role and function of the relevant reagents in the PCR reaction, understand the specific steps of the PCR reaction program.
PCR amplification of genes
Day5
pTrc99k and pMW119 plasmids extraction.
PCR amplification of genes
Day6
Enzyme digestion, connection, transformation.
Learn how to use DNA restriction endonucleases and the principle of action, understand the knowledge of DNA connection and the principle of E. coli transformation, and understand the basic concepts and operations of genetic engineering.
Construction the plasmids by recombination method and transformed into DH5α.
Day7
Pick bacteria, clone PCR identification
Learn the relevant knowledge and operation of colony PCR, understand the methods and principles of PCR identification, and learn the principles and related operations of agarose gel electrophoresis.
Culture the clonies which were identified by cloned PCR.
Day8
Escherichia coli plasmid extraction.
Sample delivery for sequencing.
Perform sequencing analysis to verify the right plasmids.
Co-transformed pTrc99k-nahR plasmids and pMW119-PsaI-EGFP plasmids into DH5α and cultured at 37℃.
Day9-10
Salicylic acid solution preparation.
Salicylic acid regulates the expression of EGFP
Day11-12
Add different concentration of salicylic acid into the co-transformed EGFP expression strains and measure the fluorescence intensity and OD600 at different time.
Use Fluorescence microscope to detect EGFP expression.
Day13-14
Add 100mM salicylic acid into the co-transformed EGFP expression strains and measure the fluorescence intensity and OD600 at different time by Spectra Max Plus 384.
Day15
Data processing. Perform relevant processing of experimental data, learn how to use related software, analyze experimental results, make preliminary judgments on experimental results, analyze the pros and cons of experimental design, find problems in the experimental process and propose optimization measures
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