About the instruments required for setting up a molecular biology laboratory
Release Date:
2025-05-16 15:12
Instruments required for setting up a molecular biology laboratory
I. Upstream Molecular Cloning
Molecular cloning technology is a core technology in molecular biology. The main purpose of this technology is to obtain a large number of copies of a certain gene or DNA fragment, thereby allowing in-depth analysis of gene structure and function, and achieving the goal of artificially modifying the genetic traits of cells and individual species.
1. Basic Technical Route of Molecular Cloning:
1) Isolation and preparation of target gene or DNA fragment;
2) Ligation of target DNA and vector in vitro;
3) Transfer of recombinant DNA molecules into host cells;
4) Screening and identification of positive recombinants;
5) Amplification of recombinants.
2. Commonly Used Instruments for Molecular Cloning:
| Instrument Name | Function |
| Air bath or water bath constant temperature shaker | Large-scale culture of cells such as bacteria or yeast |
| Centrifuge | Collection of small amounts of cells |
| Ice maker | Ice preparation |
| UV spectrophotometer | Determination of nucleic acid, protein concentration, and cell density |
| Water bath | Constant temperature treatment of samples |
| Constant temperature shaker | Constant temperature and shaking treatment of samples |
| Electrophoresis apparatus | Separation and detection of target genes |
| PCR amplifier | Amplification of target genes |
| Gel imaging analysis system | Observation and imaging of DNA fragments |
| Long-wave UV transilluminator | Excising target genes from gel |
| -80°C ultra-low freezer | Long-term storage of samples |
| Low-speed centrifuge | Collection of large amounts of cells |
| Water purifier and ultrapure water system | Preparation of deionized water and ultrapure water |
| Electroporator | Transformation or transfection of E. coli and mammalian cells |
| Digital pH meter | Measurement of sample pH |
II. Nucleic Acid Molecular Hybridization
Nucleic acid molecular hybridization technology is one of the most commonly used techniques in the field of molecular biology. Its basic principle is that two nucleic acid single strands with a certain degree of homology can form a double helix under certain conditions according to the principle of base complementarity. Due to the high specificity of nucleic acid molecular hybridization and the high sensitivity of its detection methods, it is widely used in molecular biology for screening of molecular cloning, quantitative and qualitative detection of specific gene sequences in the genome, gene expression and gene mutation analysis, and gene diagnosis of diseases, etc. According to the type of nucleic acid, it is divided into Southern blotting and Northern blotting. Commonly used instruments in nucleic acid molecular hybridization:
| Instrument Name | Function |
| Molecular hybridization oven | Hybridization of nucleic acids and other samples |
| Gel dryer | Drying the gel for long-term storage |
| Short-wave UV crosslinker | Nucleic acid immobilization |
| Electroblotting transfer apparatus | Transferring nucleic acids to membrane |
| Vacuum blotter | Transferring nucleic acids to membrane |
| DNA sequencer | DNA sequencing |
| DNA synthesizer | Primer synthesis |
| Reverse phase high pressure liquid chromatography | Purification of synthetic primers |
| Centrifugal vacuum concentrator | Concentration and drying of DNA or RNA samples |
III. Downstream Protein Expression and Purification
Whether a target gene can exert its effect can only be achieved through the expression of functional proteins. Therefore, protein expression and analysis methods have become an indispensable part of molecular biology.
1. Protein Expression
E. coli is one of the most well-known organisms in nature. Due to its advantages such as easy operation, high yield, and low cost, it has become the preferred host for protein expression. Disadvantages include: expression lacks post-translational modification, and the resulting protein may lack certain activities found in natural proteins. Yeast, as a single-celled lower eukaryote, has advantages such as being easy to cultivate, rapid reproduction, and convenient for gene manipulation, and has gradually been developed as a target gene expression system. Among them, methylotrophic yeast, as an expression system for exogenous genes, can authentically express target genes and secrete products into the culture medium, making it widely used due to features beneficial for purification. The advantages of animal cell expression systems are: the expressed products are correctly modified and generally possess natural activity, but the operation is complex, expression levels are lower, and costs are higher. All expression systems have their pros and cons and need to be selected based on experimental or production needs.
2. Instruments Required for Protein Expression
| Instrument Name | Function |
| Fermentation tank | High-density cultivation of host cells |
| Ultrasonic cell disruptor | Host cell wall disruption |
| Western blotting system | Detection of target protein |
| Liquid scintillation counter | Detection of sample radioactivity |
3. Instruments required for target protein separation and purification
Protein separation and purification is the basis for studying protein chemical composition, structure, and biological functions. In genetic engineering, downstream processing and analysis, as well as the preparation of genetically engineered products, all require proteins of high purity.
| Instrument Name | Function |
| Protein sequencer | Protein sequencing |
| Mass spectrometry sequencer | Protein sequencing |
| Nuclear Magnetic Resonance (NMR) spectrometer | Determination of protein tertiary structure |
| High-performance liquid chromatography (HPLC) instrument | Protein separation and purification |
| Ion exchange chromatograph | Protein separation and purification |
| Gel filtration chromatography column | Protein separation and purification |
| Affinity chromatograph | Protein separation and purification |
| Ultracentrifuge | Protein separation and molecular weight determination |
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