Standard procedures for the preparation of solutions commonly used in laboratories.

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keywords:solution preparation standard procedure

1, purpose

to ensure that the various solutions used in the experiment meet the experimental requirements.

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); If adopted, the implementation is published.

3, scope of application

common solutions used in XXXX research and development: 3M sodium acetate, 2N NaOH, 5M NaCL, Solution I. Solution, Solution III., LB culture base, 1M Tris-HCl, 10 ×TE Buffer, 10 × PBS Buffer, phenol/chloroform/isoquinol, 10% (W/V) SDS, 0.2 5M EDTA (pH 8.0), 50×TAEBUffer, 20 × SSC, Closed Buffer (Wester Hybrid).

4, preparation

4.1, appliances

dry>a clean 200ml bucket, two triangular bottles,balance one, 250ml, 1000ml bottles each.

4.2, preparation step

4.2.1, preparation naAC solution

(1) with balance 40.8 grams of pure NaOAC solids, placed in a 250 ml triangular burner.

(2) with 100 ml barrel accurately take 40 ml of deionized water, shake the triangle can be used to fully dissolve NaOH solids

(3) add ionized water to the solution to 100 ml

(4) autocultension, room temperature preservation.

4.2.2, the preparation of NaOH solution

(1) with a 100 ml bucket accurately take 80 ml of deionized water, placed in a 250 ml triangulation bottle.

(2) to use the balance to take 8 grams of NaOH solids slowly added to the triangular burner.

(3) shake the mixing solution. Use deionized water to build up the liquid to 100 ml.

(4) transfer the mixed solution into the glass bottle and store it at room temperature.

4.2.3, preparation of NaCL solution

(1) with balance said to take 292.2 grams of NaCL solids placed in a 1L beaker, add 800 ml of deionized water and stir to dissolve.

(2) add deionized water to the solution to 1L, the appropriate amount divided into small parts.

(3) autocultension, 4 0 C is stored.

4.2.4, Solution I. Solution

(1) take 1M Tris-HCL (ph8.0) 25ml, 1.5M EDTA (ph8.0) 20ml, 20 %Glucose (1.11M) 45ml, dH 2 O 910ml, placed in a 1L cup.

(2) high temperature autocultension, 4 0 C is stored.

(3) add 2 ml of RNaseA (20 mg / ml) to every 50 ml of Solution I. before

4.2.5, Preparation Solution II. Solution

(1) take 10%SDS 50 ml, 2N NAOH 50 ml, placed in a 500 ml beech.

(2) with sterilized water to 500 ml, well mixed.

(3) at room temperature. This solution should be kept for a period of not more than one month.

: SDS is prone to bubbles, do not stir violently.

4.2.6, Solution III. Solution

(1) Weighing KOAc 147g, CH 3 COOH 57.5g, placed in a 500 ml beo bowl.

(2) add 300 ml of deionized water and stir to dissolve.

(3) add deionized water to the solution to 500 ml.

(4) high temperature autocultension, 4 0 C is stored.

4.2.7, LB medium

(1) called T ryptone 10g, Yeast Extract 5g, NaCl 10g, placed in a 1L berrie.

(2) add about 800 ml of deionized water and stir to dissolve.

(3) drop plus 5 N NAOH (approximately 0.2 ml) to adjust the ph value to 7.0.

(4) add deionized water to the culture base to 1L.

(5) after high temperature autocultension, 4 0 C is stored.

4.2.8 preparation 1M Tris-HCl (pH7.4, 7.6, 8.0)

(1) Weighing 121.1g Tris placed in 1L berries.

(2) to add about 800 ml of deionized water. Stir thoroughly to dissolve.

(3) press the value required to add 70 ml pH7.4, 60 pH7.6, 42 ml pH8.0 hydrochloric acid regulation.

(4) to the solution to 1L.

at room temperature after (5) high temperature autocultension.

note: the pH is adjusted after cooling the solution to room temperature, as the pH of the Tris solution varies greatly with the temperature, which decreases by about 0.03 units for every 1 degree C increase in temperature.

4.2.9, 1L 10 ×TEBuffer (pH7.4, 7.6, 8.0) 100mM Tris-HCl, 10mM EDTA

(PH7.6, 8.0) 1) Measure solution 1M Tris-HClBuffer (pH7.4,, 7.6, 8.0) 100ml, 500mM EDTA (pH 8.0) 20ml, placed in a berries.

(2) add 800 ml of deionized water to the beech and mix evenly.

high temperature and autocultension after the solution has been dissolved to 1L at the same time (3).

(4) room temperature save.

4.3.10, 1L 10 × PBS Buffer 1370mM NaCl, 27mM KCl, 10mM Na 2 HPO 4, 20mMM KH 2 PO 4 .

(1) weighing NaCl 80g, KCl 2, Na 2 HPO 4 14.2g, KH 2 PO 4, 2.7g in a 1L beech.

(2) add 800 ml of deionized water to the beech and stir to dissolve.

(3) drop plus HCl to adjust the pH to 7.4, and then add deionized water to the solution to 1L.

(4) high temperature autocultension, room temperature preservation.

note that there are no two-priced cations in the PBSBuffer above, and 1m CaCl 2 and 0.5mMM MgCl 2 can be added to the formula.

4.3.11 Preparation of phenol/chloroform/isoquinol (25:24:1)

(1) Description: removal from nucleic acidsprotein common use of phenol/chloroform/isoprenol (25:24:1). Chloroform denatures proteins and helps separate the liquid phase from the organic orientation, while isoquinol helps eliminate bubbles that appear in the drawer process.

(2) configuration method: the balance of phenol and the same volume of chloroform / isosterol (24:1) evenly mixed and after moving into the brown glass bottle 4 degrees C to save.

4.3.12 preparation 10% (W/V) SDS 100ml

(1) weighing 10g high-weight SDS placed in 100-200ml berries, adding about 800ml of deionized water, 68C heatingdissolved.

(2) drops plus a few drops of thick hydrochloric acid to adjust the pH to 7.2.

(3) to 100 ml, store at room temperature.

4.3.13, preparation 1L 0.5M EDTA (pH 8.0)

(1) 186.1gNa 2 EDTA.2H 2 O, placed in a 1L berred cup.

(2) to add 800ml of deionized water and stir well.

(3) adjust the pH to 8.0 (about 20g NOH) with NaOH Note: pH to 8.0 can not be fully dissolved.

(4) plus deionized water to the solution to 1L.

high temperature autocultension after the temperature (5) was divided into small points in moderation.

at room temperature (6).

4.3.14 preparation 1L 50 ×TAE BUffer (pH8.5) 2M Tr.