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(12) why does the reagent evaporate or even dry in the PCR e

时间:2018-05-02 来源:EASTWIN浏览次数:106

             Problem diagnosis             
 
Dongdong: why does reagent in PCR experiment evaporate? Even steamed dry? What should I do?

Long Long: evaporation of the reagent is one of the common problems in PCR experiment. There are many factors that affect the evaporation of the reagent.

Evaporation is the reagent in the process of PCR experiment, the reaction liquid is vaporized at high temperature and condenses on the wall or top of the tube, forming water mist or water droplets, or escaping directly from the gap of the tube cover or the film covering, resulting in the change of the concentration of the reaction liquid, the decrease of the amount of the reaction liquid, and some even steaming, resulting in the ineffectiveness of the experiment.

In the PCR experiment, we generally use the heating cover to prevent the condensation of the reagent steam. The heat cover temperature is generally higher than the reaction liquid, in order to prevent the condensation of the steam.

For steam leakage, most people think that the pressure of hot cover is not enough. The pressure of hot cover is very important, but this is only one aspect. In fact, consumables are also crucial.

The quality of the test tube, the mouth of the pipe and the lid are smooth, the sealing of the box lid is good, and it is not easy to leak. Poor quality consumables, uneven pipes, radial pattern, easy to cause leakage, or even dry.

For the PCR reaction of small system (5-10ul), 96 hole plates are mostly used for experiments. 96 hole plates are usually sealed by plastic film, and are also covered with silicone rubber. If it is covered with plastic film, the material, thickness, hardness, film temperature, pressure and smoothness of the plastic film will all be affected. Generally choose soft, thick film is better. In order to increase the sealing pressure of the nozzle, a soft, 96 raised silicone pad can also be added to the hot cover.

In addition, the parameter setting of the PCR program is also important, and the reaction system that is closest to the actual reaction system is selected. For example, the actual reaction system is 5ul. If 25ul and even 50ul are selected when the program is set up, the reagents are overwashed at 95 degrees centigrade, and the time of dynamic temperature balance will be prolonged. This actually increases the temperature of the reagent (it may reach 98 degrees C), and prolongs the time to stay at the high temperature, greatly increasing the risk of reagents leaking, and even making the reagents completely evaporated (Zheng Gan).


If you have other problems, you can also contact our after-sale Engineer Yang Gong anytime, Tel: 18051112172. Suzhou Dongsheng Xingye Scientific Instrument Co., Ltd. provides you with the most reliable service guarantee!!!
 

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