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  • pcr实验室设计要注意什么?
    来源:http://www.jnyhrs.com 浏览: 发布日期:2021-12-30
    一、PCR实验室布局
    1、 PCR laboratory layout
    PCR实验室原则上为试剂准备区,样品制备区,扩增区和扩增产物分析区四个单独的工作区域,并设有专用走廊,各工作区域应设缓冲间,工作区与缓冲间宜安装连锁装置。不同功能的工作区应是分隔独立的,各工作区有明显的标志,不能直通,如果紧密连接,需安装物品传递窗。前两区为扩增前区,后两区为扩增后区,即从:试剂准备区一样品制备区一 PCR 扩增室一产物分析室 ,不得逆向流动。实验室的气流也应从扩增前区流向扩增后区,不得逆向流动。各工作区顶部应该安装紫外灯,紫外灯波长为254mm,每20m一支40W的紫外灯。
    In principle, PCR laboratory is divided into four separate working areas: reagent preparation area, sample preparation area, amplification area and amplification product analysis area, with special corridors. Buffer rooms shall be set in each working area, and interlocking devices shall be installed between the working area and buffer room. Work areas with different functions shall be separated and independent. Each work area shall have obvious signs and cannot be connected directly. If they are closely connected, article transfer windows shall be installed. The first two areas are pre amplification areas, and the last two areas are post amplification areas, i.e. from reagent preparation area - sample preparation area - PCR amplification room - product analysis room. Reverse flow is not allowed. The air flow in the laboratory shall also flow from the pre amplification area to the post amplification area, and shall not flow in reverse. Ultraviolet lamps shall be installed at the top of each working area. The wavelength of ultraviolet lamps is 254MM, and one 40W ultraviolet lamp shall be installed every 20m.
    二、实验室各区功能及主要设备
    2、 Functions and main equipment of each area of the laboratory
    1.试剂准备区:主要进行试剂的制备,分装和主反应混合液的制备。试剂盒用于样品制作的材料应直接运送至该区域。试剂原材料必须贮存在本区内,并在本区内制备成所需的试剂。本区主要设备有天平,冰箱,离心机,加样器,振荡器等。对于气流压力的控制,本区并没有严格的要求, 一般为+10Pa。
    1. Reagent preparation area: mainly for reagent preparation, sub packaging and preparation of main reaction mixture. The materials of the kit for sample preparation shall be transported directly to the area. Reagent raw materials must be stored in this area and prepared into required reagents in this area. The main equipment in this area includes balance, refrigerator, centrifuge, sampler, oscillator, etc. There are no strict requirements for the control of air flow pressure in this area, which is generally + 10Pa.
    实验室装修
    2.样品制备区:主要进行样品的保存,核酸提取,贮存及其加入至扩增反应管和测定DNA的合成。本区主要设备有冰箱,生物安全柜,离心机,加样器,振荡器等。本区的压力梯度要求为:相对于邻近区域为正压,以避免从邻近区进入本区的气溶胶污染, 一般为+10Pa.
    2. Sample preparation area: it is mainly used for sample preservation, nucleic acid extraction, storage, addition to amplification reaction tube and DNA synthesis. The main equipment in this area includes refrigerator, biosafety cabinet, centrifuge, sampler, oscillator, etc. The pressure gradient in this area is required to be positive pressure relative to the adjacent area to avoid aerosol pollution entering this area from the adjacent area, which is generally + 10Pa
    3.扩增区:主要进行DNA扩增。此外,已制备的DNA模板(来.自样品制备区)的加入和主反应混合液(来自试剂准备区)制备成反应混合液等也可在本区内进行。本区主要设备有:扩增仪,冰箱,离心机,加样器等。本区的压力梯度要求为:相对于邻近区域为负压,以避免气溶从本区漏出一般为一25Pa。
    3. Amplification area: mainly for DNA amplification. In addition, the addition of the prepared DNA template (from the sample preparation area) and the preparation of the main reaction mixture (from the reagent preparation area) into the reaction mixture can also be carried out in this area. The main equipment in this area include: amplification instrument, refrigerator, centrifuge, sampler, etc. The pressure gradient in this area is required to be negative pressure relative to the adjacent area to avoid gas solution leakage from this area, which is generally a 25 PA.
    4.扩增产物分析区:扩增产物的测定。本区主要设备有酶标仪,洗板机、加样器和水浴箱等。本区的压力梯度的要求为:相对于邻近区域为负压,一般为一25Pa,以避免扩增产物从本区扩散至其它区域。
    4. Amplification product analysis area: Determination of amplification products. The main equipment in this area include enzyme labeling instrument, plate washer, sampler and water bath box. The pressure gradient in this area is required to be negative pressure relative to the adjacent area, generally 25 PA, so as to avoid the diffusion of amplification products from this area to other areas.
    三、PCR实验室通风系统及压力控制
    3、 Ventilation system and pressure control of PCR Laboratory
    各区域之间应具备单间的实验工艺流,物流,人流与气流,形成单向流程的保护屏障, 避免实验室之间的相互干扰,防止气溶胶扩散对实验过程造成污染。PCR实验室并没有严格的净化要求,但是为避免各个实验区域交叉污染的可能性,宜采用全送全排的气流组织形式,严格控制送,排风的比例以保证个实验区的压力要求。
    There shall be single room experimental process flow, logistics, people flow and air flow between each area to form a protective barrier of one-way flow, avoid mutual interference between laboratories and prevent aerosol diffusion from polluting the experimental process. PCR laboratory has no strict purification requirements, but in order to avoid the possibility of cross pollution in each experimental area, it is appropriate to adopt the air distribution form of full supply and full exhaust, and strictly control the proportion of supply and exhaust to ensure the pressure requirements of each experimental area.
    四、PCR实验室装饰
    4、 PCR laboratory decoration
    实验室围护结构应牢固, 气密性好;所有阴阳角宜采用圆弧过度;墙体内壁光洁,不积尘,耐腐蚀,易清洗消毒;地面使用PVC卷材或环氧树脂自流坪,材料应满足无缝隙,无渗漏,光洁,耐腐蚀的要求。
    The enclosure structure of the laboratory shall be firm with good air tightness; Arc transition shall be adopted for all internal and external corners; The inner wall of the wall is smooth, dust free, corrosion resistant and easy to clean and disinfect; PVC coiled material or epoxy resin self flow floor shall be used on the ground, and the material shall meet the requirements of no gap, no leakage, smoothness and corrosion resistance.