A summary of buffers is commonly used in the laboratory.

1, 1M Tris-HCl (pH7.4, 7.6, 8.0)
□ concentration 1 M Tris-HCl
□ preparation 1L
□ configuration method 1. Weighing 121.1gTris placed in 1L
berries
.
2. Add about 800mL of deionized water and stir to dissolve.
3. Add the pH required for hydrochloric acid regulation by adding the table below.
pH thick HCl
7.4 about 70mL
7.6 about 60mL
8.0 about 42mL
4. Dissolve to 1L.
5. After high temperature autocultension, room temperature is preserved.
: The solution should be cooled to room temperature before setting the pH, because the pH of the Tris solution varies greatly with the temperature, and the pH of the solution decreases by about 0.03 units for every 1 degree C increase in temperature.
2, 1.5 M Tris-HCl (pH8.8)
□ ingredient concentration 1.5 M Tris-HCl
□ preparation 1L
□ configuration method 1. 181.7gTris is placed in a 1L berred.
2. Add about 800mL of deionized water and stir to dissolve.
3. Adjust the pH to 8.8 with hydrochloric acid.
4. The solution is fixed to 1L.
5. After high temperature autocultension, room temperature is preserved.
note: The solution should be cooled to room temperature before the pH is adjusted, because the pH of the Tris solution varies greatly with the temperature, and the pH of the solution decreases by about 0.03 units for every 1 degree C increase in temperature.
3, 10×TE (pH 7.4, 7.6, 8.0)
□ ingredient concentration 100 mM Tris-HCl, 10 mM
EDTA □ preparation 1L
□ configuration method 1. quantity of the following solution, placed in a 1L berries.
1 M Tris-HCl Buffer (pH7.4, 7.6, 8.0) 100mL
500 mM EDTA (pH8.0) 20mL
2. Add approximately 800mL of deionized water to the beech and mix evenly.
3. After the solution is set to 1L, high temperature and high pressure sterilization.
4. Store at room temperature.
4, 3 M sodium acetate (pH5.2)
□ group concentration 3 M sodium acetate
□ ingredients 100mL
□ configuration method 1. 40.8g NaOAc-3H2O is placed in a 100-200mL beo bowl, add about 40mL of deionized water stirring solution.
2. Add ice acetic acid to adjust the pH to 5.2.
3. Add deionized water to the solution to 100mL.
4. After high temperature and autocultension, room temperature is preserved.
5, PBS Buffer . □ concentration 137 mM NaCl, 2.7mM KCl, 10 mM Na2HPO4, 2 mM KH2PO4
□ dosing 1L
□ configuration method 1. Weighing the following
reagents
, placed in a 1L berring cup.
NaCl8 g
. KCl0.2g
Na2HPO41.42 g
KH2PO40.27g
2. Add about 800 mL of deionized water to the beech and stir to dissolve.
3. Drip HCl adjusts the pH to 7.4, and then adds deionized water to the solution to 1L.
4. After high temperature and autocultension, room temperature is preserved.
note: There are no two-priced cations in the PBS Buffer above, and 1mM CaCl2 and 0.5 mM MgCl2 can be added to the formula if required.
6, 10 M ammonium acetate . □ concentration of 10 M ammonium acetate
□ the preparation amount of 100mL
□ configuration method 1. Weighing 77.1g ammonium acetate is placed in a 100 to 200 mL berries and dissolved by stirring and dissolving about 30 mL of deionized water.
2. Add deionized water to the solution to 100mL.
3. Use a 0.22 m membrane
filter
sterilization.
4. Seal the mouth of the bottle at room temperature.
: ammonium acetate is easily decomposed by heat, so it can not be sterilized at high temperature and high pressure.
7, Tris-HCl Balanced Phenol
□ Configuration Method
1. Using raw materials: Most commercially available liquefied phenols are bright and colorless and can be used in
molecular biology
experiments without re-distillation. However, some lily phenols are pink or yellow and should be avoided. At the same time, the use of crystalline phenol should also be avoided, crystalline phenol must be re-distilled at 160 degrees C to remove oxidizing products such as niobium, these oxidation products can cause the break of the phosphate bonds or lead to RNA and
DNA
cross-linking. Therefore, the quality of phenol is very important for dna, RNA extraction, we recommend the use of high-quality phenol for molecular biology experiments.
: Phenol is highly corrosive and can cause serious burns, wear
gloves
protective
etc. All operations should be carried out in the ventilation cabinet, with a large amount of water cleaning applied to the skin areas in contact with phenol, and washed with soap and water, avoid ethanol.
3. Phenol balance: Since DNA is distributed in the organic phase under acidic pH conditions, phenol must be balanced to a pH of more than 7.8 before use, and the phenol balance operation method is as follows:
(1) LPHEN should be stored at -20 degrees C, at which point the phenol is crystalline. The phenol removed from the freezer is first placed at room temperature to reach room temperature, and then dissolved fully in a 68-degree water bath.
(2) add hydroxyquine (8-Quinolinol) to the final concentration of 0.1%. The compound is a reducing agent, an incomplete inhibitor of RNA enzymes, and a weak chelating agent of metal ions, also because it is yellow. It helps to identify organic phases easily.
(3) add 1M Tris-HCl (pH8.0) of the same volume and stir for 15 minutes using a magnetic agitator, set aside to allow it to be fully layered, and remove the upper water phase.
(4) repeat the procedure (3).
(5) add 0.1M Tris-HCl (pH8.0) of the same volume, stir for 15 minutes using a magnetic agitator, set aside to allow it to be fully layered, and remove the upper water phase.
(6) repeat the operating procedure (5), slightly residual part of the upper water phase.
(7) use pH
paper
confirm that the pH of the organic phase is greater than 7.8.
(8) to place phenol in a brown glass bottle for 4 degrees C to preserve.
.