Method for one-step dry powdering molecular biological reagent
A technology of biological reagents and sugar substances, which is applied in the field of molecular biology and can solve the problems of lack of protective agents
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[0055] In another preferred example, the preparation method of the dry powdering reagent of the present invention comprises the following steps:
[0056] (1) Prepare a protein-containing solution, which contains protein and optional other components (such as the components of the reaction system) and a certain concentration of vacuum drying protective agent (such as polysucrose) solution;
[0057] (2) Mix the protein solution and the lyoprotectant solution in a certain proportion to form a reaction mixture, and then distribute the reaction mixture according to a certain volume (the purpose of the packaging is to add a certain volume of liquid substrate during the experiment to carry out the reaction) ;
[0058] (4) Vacuum drying or freeze-drying to obtain dry powdered reagents. Usually, it can be frozen at about -80°C first, and then vacuum dried. Of course, you can also vacuum dry directly without going through the freezing step ;)
[0059] The dry powdering reagent of the...
Embodiment 1
[0070] Dry Powdered Enzyme Reagents
[0071] Add polysucrose to 1 μl of purchased M-MLV enzyme containing 200U at a weight ratio of 0.1:1, 1:1, and 1-0.1, store at 4°C for 72 hours, and then determine by measuring the relative fluorescence intensity in the amplification reaction enzyme activity. Controls were M-MLV enzymes stored at -20°C and 4°C for 72 hours without adding Ficoll.
[0072] relative activity
Ficoll-free M-MLV enzyme stored at -20°C for 72 hours
100％
Ficoll-free M-MLV enzyme stored at 4°C for 72 hours
10％
Store at 4°C for 72 hours, add 0.1:1 weight ratio of Ficoll M-MLV enzyme
＞90％
Store at 4°C for 72 hours, add 1:1 weight ratio of Ficoll M-MLV enzyme
＞85％
Store at 4°C for 72 hours, add 1:0.1 weight ratio of Ficoll M-MLV enzyme
80％
[0073] This shows that the addition of polysucrose can significantly and effectively prolong the activity of the enzyme preserved at room temperatu...
Embodiment 2
[0076] Dry powdered RNA reverse transcription (reverse transcription; RT) reagent
[0077] A RT-PCR Pre-Mix formulation, the specific composition of which is shown in Table 1. The mix contains everything needed to perform an RT reaction except the RT primers and RNA. After the dry powder reagent is made, just add an appropriate volume of sample RNA to carry out RT-PCR reaction.
[0078] Components (initial concentration)
[0079] After preparing the above reagents, vacuum-dry them directly, or freeze them at -80°C and then vacuum-dry them to make RT-PCR reaction dry powder reagents.
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