Construction scheme of molecular biology laboratory
Construction scheme of molecular biology laboratory
As the upstream technology of genetic engineering, molecular biology, the results and accuracy of its experiments will determine all the downstream steps and the final experimental results. Therefore, it is very important to build a complete molecular biology laboratory. Let us take a look at what instruments are needed to build a molecular biology laboratory.
1. Refrigerator: According to the storage needs of medicines, reagents and various biological agents, refrigerators with different temperature control levels must be equipped. The most commonly used ones are: 4 ℃, -20 ℃, -80 ℃ refrigerators. 4 ℃ is suitable for storing certain solutions, reagents, medicines, etc. -20 ℃ is suitable for certain reagents, medicines, enzymes, serum, prepared antibiotics, DNA, protein samples, etc. -80 ℃ is suitable for the preservation of some long-term cryopreserved samples, E. coli strains, purified samples, special low-temperature processed digestive juice, etc. Chromatography freezer at 0-10 ℃ is suitable for electrophoresis, chromatography, dialysis and other experiments under low temperature conditions.
2. Incubator: 37 ° C incubator is used for bacterial plate cultivation and molecular biology experiments.
3. Constant temperature culture shaker: used for the amplification of E. coli and other biological engineering strains.
4. Water bath: used for heat preservation and various experiments.
25-100 ℃ water bath shaker can be used for the thermal insulation of molecular hybridization experiments and various biochemical enzyme reactions.
The low-temperature water bath can be used for experiments such as the connection of plasmids and gene fragments in molecular biology and the heat shock of E. coli competent at 42 degrees.
5. Oven: It is mainly used for drying experimental utensils, some need higher temperature, some need lower temperature. Experimental equipment for RNA needs to be dried in an oven at 250 ° C. Some plastic appliances can only be dried in an oven at 42-45 ° C.
6. Ultra-pure water machine: With the rapid development of molecular biology, many experiments have higher and higher requirements for water purity. Tap water, distilled water, ion-exchanged water, reverse osmosis pure water are used as water supply, and the magnet is coupled with gear pump Circulate water. Used for PCR, PCR amino acid analysis, DNA sequencing, enzyme reaction, tissue and cell culture, etc.
7. Steam sterilization pot: Most reagents used in molecular biology, and laboratory equipment should be strictly sterilized. It can be used to disinfect small batches at any time. A large number of experimental items, reagents, and culture media can be disinfected on a large scale and regularly.
8. Filter membrane: Reagents that are not resistant to high temperature and high pressure can be used for bacteria.
9. Various balances: platform scales, precision electronic analytical balances, used to accurately weigh all kinds of reagents.
10. Measurement of liquid volume: measuring cylinder, pipette, micro pipette, graduated test tube, beaker, conical flask, etc.
11. pH meter: an instrument that measures the direct potential of H + in a solution, mainly through a pair of electrodes, which generates different electromotive forces in different pH solutions and expresses it with a pH value;
pH test paper: only suitable for rough estimation of the pH value of the culture solution, phenol saturated solution, buffer solution or other reagent solutions. However, the preparation of most reagents requires strict pH values, requiring a pH meter with high accuracy (two decimal places).
12. Spectrophotometer: Optical density and spectrophotometer is an instrument that uses the absorption spectrum of a substance in the visible and ultraviolet regions to identify the substance and its content. It is composed of light source, monochromator, absorption cell, receiver, measuring instrument or display screen. The OD value is one of the convenient indicators for quantification of solutes in many solutions. The monochromatic light generated can be used to determine the absorption value of a solution for the monochromatic light, and it can be used for preliminary judgment of nucleic acid solution quantification and purity. The OD value can also be used as a detection index for bacterial concentration.
13. High-speed centrifuge: The maximum speed is 25000 r / m and the maximum centrifugal force is 89000g. There are two kinds of freezing and normal temperature, mostly used for preparation and hand collection of microorganisms, cell debris, cells, large organelles, ammonium sulfate precipitates and immunoprecipitates.
14. Ultra-clean workbench: There are ultraviolet lamps, lighting lamps, alcohol lamp flame, 75% ethanol and other sterilization equipment, which is a device that provides local cleanliness. The principle is that the blower drives the air, passes through the low- and medium-efficiency filters, and passes through the work surface to make the experimental operation area a sterile environment.
15. Electrophoresis system: Electrophoresis technology is one of the tools for detecting and identifying the purity and content of various biological macromolecules and describing their characteristics, and even separating, purifying, recovering and concentrating samples.
The electrophoresis system is divided into a power supply and an electrophoresis tank. The power supply needs to pass a steady flow through the voltage regulator, which can provide stable DC power and output a stable voltage; horizontal electrophoresis tank: generally divided into mini electrophoresis tank and large horizontal electrophoresis groove
16. PCR instrument: Polymerase Chain Reaction instrument, also known as DNA thermal cycler, gene amplification instrument, it is a pair of oligonucleotide primers bound to both sides of the target sequence on the positive and negative DNA strands, thereby enzymatically synthesizing copies Millions of times the target DNA fragment, each cycle of it includes three processes: DNA denaturation at three different temperatures, primer renaturation, and DNA polymerase-catalyzed extension reaction. It should be noted that some laboratories may also need a gradient PCR instrument or a practical quantitative fluorescence PCR instrument to perform some special molecular biology experiments.
17. Gel imaging analysis system: Biotek UltraPowerTM visible light gel projector is a good choice, the price is less than seven thousand, the detection sensitivity can reach pg level (UV and EB can only reach 5ng), and UV and EB are not needed Safer, easy to operate and take pictures.
18. Other equipment
1. Microwave oven: convenient for rapid heating and constant temperature heating of some solutions, preparation and melting of electrophoresis agarose gel.
2. Ice machine: used in the low temperature environment required for the experimental operation of most nucleic acids and proteins to reduce the hydrolysis of nucleases or proteinases.
3. Chromatography device: (chromatographic separation) is an effective physical method for separating multi-component mixtures.
Vacuum imprinting system, DNA synthesis / sequencer: These are all necessary instruments for in-depth study of nucleic acids.
4. Magnetic stirrer: multi-angle rotating mixer, rapid oscillation mixer: used for mixing instruments.
5. Tissue homogenizer: Ultrasonic tissue and cell disrupter, used for sample separation and purification experiments.
6. Fume hood: Many solvents can escape poisonous gas, a necessary cabinet, and radiation test must also be shielded by organic glass.
7. Glass distiller, electric heating cap, transformer: used for distillation of phenol and other organic solvents.
8. Tip head, Eppendorf tube:
The tip of the microtube pipette (aspirating tip) and the Eppendorf tube (microcentrifuge tube) can be washed, siliconized and sterilized and used repeatedly. For some strict experiments, such as RNA extraction and preservation, new sterile tip and Eppendorf tube should be used. In addition, common specifications of centrifuge tubes (1000ml, 500ml, 250ml, 50ml, 7ml, etc.) and 96-well, 24-well, 12-well, 6-well cell culture plastic plates, etc. should also be prepared.
9. Small equipment and utensils:
Timers, filter membranes, plastic wraps, protective glasses duckbill tweezers, conventional glass or plastic utensils (including plates, test tubes, beakers, measuring flasks, reagent separatory funnels, and reagents stored in dark place should use brown reagent bottles, such as saturated Phenol, mercaptoethanol, etc.), marker pen, various gloves PE, latex, household, acid-proof, etc.)
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