1. 30% acrylamide solution
ã€Preparation method】 Dissolve 29g acrylamide and 1g N, N-methylenebisacrylamide in water with a total volume of 60ml. Heat to 37 ℃ to dissolve, add water to make the final volume 100ml. Filter and sterilize with a filter (0.45μm pore size), verify that the pH value of the solution should not be greater than 7.0, and store in a brown bottle at room temperature.
[Note] Acrylamide has strong neurotoxicity and can be absorbed through the skin, and its effect is cumulative. Gloves and masks should be worn when weighing acrylamide and methylenebisacrylamide. Polyacrylamide can be considered non-toxic, but it should be handled with caution, as it may also contain small amounts of unpolymerized material.
Some less expensive acrylamide and bisacrylamide usually contain some metal ions. Add about 0.2 volume of single bed mixed resin (MB-1 Mallinckrodt) to the acrylamide stock solution, stir overnight, and then filter with Whatman No. 1 filter paper for purification Of it.
During storage, acrylamide and bisacrylamide are slowly converted to acryl and bisacrylic acid.
2. 40% acrylamide
[Preparation method] Dissolve 380g of acrylamide (DNA sequencing grade) and 20g of N, N-methylenebisacrylamide in 600ml of distilled water. Continue to process according to the above method of preparing 30% acrylamide solution, but after heating and dissolving, make up with distilled water to a final volume of 1L.
[Note] See the above instructions for preparing 30% acrylamide, 40% acrylamide solution for DNA sequence determination.
3. Actinomycin D solution
[Preparation method] Dissolve 20 mg of actinomycin D in 4 ml of 100% ethanol, dilute the storage solution 1:10, and use 100% ethanol as a blank control to read the OD440 value. The molar digestibility coefficient of the actinomycin D (molecular weight 1255) pure product in aqueous solution is 21,900, so the absorbance of the 1mg / ml solution of actinomycin D at 440 nm is 0.182 It should be placed in a test tube wrapped with foil and stored at -20 ° C.
[Note] Actinomycin D is a teratogen and a carcinogen. When preparing this solution, you must wear gloves and operate in a fume hood, not open on the experimental table. skin.
Actinomycin D products provided by pharmaceutical companies for therapeutic purposes often contain additives such as sugar or salt. As long as the concentration of actinomycin D is determined by measuring the light absorption of the storage solution at a wavelength of 440 nm, such products can be used to suppress their own guiding effect
4. 0.1mol / L adenosine triphosphate (ATP) solution
[Preparation method] Dissolve 60mg ATP in 0.8ml water, adjust the pH value to 7.0 with 0.1mol / L NaOH, dilute 1ml with distilled water, and store in small portions at -70 ℃
5. 10mol / L acetic acid solution
[Preparation method] Dissolve 770g of acetic acid in 800ml of water, add water to make up to 1L, then filter and sterilize.
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