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Standard procedures for the preparation of common solutions in laboratories

1. Purpose
Ensure that the various solutions used in the experiment meet the requirements of the experiment.
2. The SOP change procedure
Changes to this document can be proposed by any staff member who uses the document and reported to the person in charge for decision. If passed, it will be announced and implemented.
3. Scope of application
Common solutions: 3M sodium acetate, 2N NaOH, 5M NaCL, Solution Ⅰ, Solution Ⅱ Solution, Solution Ⅲ, LB medium, 1M Tris-HCl, 10 × TE Buffer, 10 × PBS Buffer, phenol/chloroform/isoamyl alcohol, 10% (W/V) SDS, 0.5M EDTA (pH 8.0), 50 × TAEBUffer, 20 × SSC, blocking buffer (Wester hybridization).
4. Match
4.1 、 Utensils
One dry and clean 200ml measuring barrel, two Erlenmeyer flasks, one balance, one 250ml and one 1000ml wide-mouth flask.
4.2 、 Preparation steps
4.2.1 、 Prepare NaAC solution
⑴Weigh 40.8 grams of analytically pure NaOAC solid with a balance and place it in a 250ml Erlenmeyer flask.
⑵Accurately take 40ml deionized water in a 100ml measuring bucket, shake the Erlenmeyer flask to fully dissolve the NaOH solid
⑶ Add deionized water to dilute the solution to 100ml
⑷After autoclaving, store at room temperature.
4.2.2 Preparation of NaOH solution
⑴Accurately take 80ml of deionized water in a 100ml measuring bucket and place it in a 250ml Erlenmeyer flask.
(2) Weigh 8 grams of NaOH solid with a balance and slowly add it to the Erlenmeyer flask.
⑶ Shake and mix the solution. The volume of the solution was adjusted to 100ml with deionized water.
⑷ Transfer the mixed solution into a glass bottle and store at room temperature.
4.2.3 、 Preparation of NaCL solution
⑴Weigh 292.2g of NaCL solid with a balance and place it in a 1L beaker, add 800ml of deionized water and stir to dissolve.1000 Beaker,50 Ml Beaker,Beaker With Spout,Lab Beaker 5000ml
⑵Add deionized water to dilute the solution to 1L, and divide the appropriate amount into small portions.
⑶ After autoclaving, store at 40°C.
4.2.4. Prepare Solution Ⅰ
(1) Measure 25ml of 1M Tris-HCL (ph8.0), 20ml of 1.5M EDTA (ph8.0), 45ml of 20%Glucose (1.11M), 910ml of dH 2 O, and place them in a 1L beaker.
(2) After autoclaving, store at 40°C.
(3) Add 2 ml of RNaseA (20mg/ml) for every 50 ml of Solution Ⅰ before use.
4.2.5. Prepare Solution Ⅱ
(1) Measure 50 ml of 10% SDS, 50 ml of 2N NAOH, and place in a 500 ml beaker.1000 Beaker,50 Ml Beaker,Beaker With Spout,Lab Beaker 5000ml
(2) Add sterile water to make the volume to 500 ml, and mix thoroughly.
(3) Store at room temperature. It is best not to keep this solution for more than one month.
Note: SDS is easy to produce bubbles, do not stir vigorously.
4.2.6. Prepare Solution Ⅲ
(1) Weigh 147g of KOAc and 57.5g of CH 3 COOH and place them in a 500 ml beaker.
(2) Add 300 ml of deionized water and stir to dissolve.
(3) Add deionized water to dilute the solution to 500 ml.
(4) After autoclaving, store at 40°C.
4.2.7 Preparation of LB medium
(1) Weigh T ryptone 10g, Yeast Extract 5g, and NaCl 10g, and place them in a 1L beaker.
(2) Add about 800 ml of deionized water, stir well to dissolve.
(3) Add 5 N NAOH (about 0.2ml) dropwise and adjust the pH to 7.0.
(4) Add deionized water to dilute the medium to 1L.
(5) After autoclaving, store at 40°C.
4.2.8 Preparation of 1M Tris-HCl (pH 7.4, 7.6, 8.0)
(1) Weigh 121.1g Tris in a 1L beaker.
(2) Add about 800ml of deionized water. Stir well to dissolve.
(3) Press to add 70ml pH7.4, 60 pH7.6, 42ml pH8.0 concentrated hydrochloric acid to adjust the required value.
(4) Dilute the solution to 1L.
(5) Store at room temperature after autoclaving.
Note: Adjust the pH value after cooling the solution to room temperature, because the pH value of the Tris solution varies greatly with temperature, and the value of the solution decreases by approximately 0.03 units for every 1°C increase in temperature.
4.2.9 Prepare 1L 10×TEBuffer (pH7.4,7.6,8.0) 100mM Tris-HCl, 10mM EDTA
(1) Measure 100 ml of 1M Tris-HClBuffer (pH 7.4, 7.6, 8.0) and 20 ml of 500 mM EDTA (pH 8.0) in a beaker.
(2) Add 800ml of deionized water to the beaker and mix evenly.
(3) After dissolving the solution to 1L, sterilize it under high temperature and high pressure.
(4) Store at room temperature.
4.3.10. Prepare 1L 10 × PBS Buffer 1370mM NaCl, 27mM KCl, 10mM Na 2 HPO 4, 20mM KH 2 PO 4.
(1) Weigh 80g NaCl, KCl 2, Na 2 HPO 4 14.2g, KH 2 PO 4, 2.7g and place them in a 1L beaker.
(2) Add 800ml of deionized water to the beaker and stir to dissolve it.1000 Beaker,50 Ml Beaker,Beaker With Spout,Lab Beaker 5000ml
(3) Add HCl dropwise to adjust the pH to 7.4, then add deionized water to dilute the solution to 1L.
(4) Sterilize at high temperature and high pressure, store at room temperature.
Note: There are no divalent cations in the above PBSBuffer. If necessary, 1mM CaCl 2 and 0.5mM MgCl 2 can be added to the formula.
4.3.11 Preparation of phenol/chloroform/isoamyl alcohol (25:24:1)
(1) Description: Phenol/chloroform/isoamyl alcohol (25:24:1) is often used to remove protein from nucleic acids. Chloroform denatures proteins and helps to separate the liquid phase from the organic phase, while isoamyl alcohol helps to eliminate air bubbles in the drawer process.
(2) Configuration method: After the balanced phenol and equal volume of chloroform/isoamyl alcohol (24:1) are uniformly mixed, they are transferred to a brown glass bottle and stored at 4°C.
4.3.12 Prepare 10% (W/V) SDS 100ml
(1) Weigh 10g of high-severity SDS in a 100-200ml beaker, add about 800ml of deionized water, and heat to dissolve at 68°C.
(2) Add a few drops of concentrated hydrochloric acid to adjust the pH to 7.2.
(3) After diluting the solution to 100ml, store at room temperature.
4.3.13 、 Prepare 1L 0.5M EDTA (pH 8.0)
(1) Weigh 186.1g of Na 2 EDTA.2H 2 O and place it in a 1L beaker.
(2) Add 800ml deionized water and stir well.
(3) Adjust the pH value to 8.0 with NaOH (about 20g NOH). Note: It can be completely dissolved when the pH value reaches 8.0.
(4) Add deionized water to dilute the solution to 1L.
(5) After the appropriate amount is divided into small parts, it is sterilized by high temperature and high pressure.
(6) Store at room temperature.
4.3.14 Prepare 1L 50×TAE BUffer (pH8.5) 2M Tris acetic acid, 100mM EDTA
(1) Weigh 242g of reagent Tris and 37.2g of Na 2 EDTA.2H 2 O and place them in a 1L beaker.
(2) Add about 800ml of deionized water to the beaker, stir well and dissolve.
(3) Add 57.1ml of acetic acid and stir well.
(4) After making the volume of deionized water to 1L, store at room temperature.
4.3.15 、 Prepare 1L 20×SSC 3.0M NaCl, 03M sodium citrate
(1) Weigh 175.3g of reagent NaCl and 88.2g of sodium citrate. 2H 2 O, and place them in a 1L beaker.
(2) Add about 800ml of deionized water to the beaker, stir well to dissolve.1000 Beaker,50 Ml Beaker,Beaker With Spout,Lab Beaker 5000ml
(3) Add 14 N HCl dropwise, adjust the pH to 7.0, add deionized water to make the volume up to 1L.
(4) Store at room temperature after autoclaving.
4.3.16 Prepare 100ml pre-hybridization solution/hybridization solution (for DNA hybridization), 6×SSC (or SSPE), 5×Denhardts 0.5%(WV) SDS, 100μg/ml Salmon DNA
(1) Weigh 30ml 20×SSC (or SSPE) reagents, 10ml 50×Denhardts, 5ml 10% SDS, 1ml 10μg/ml Salmon DNA, 54m dl2H 2 O, and place them in a 200ml beaker.
(2) After mixing thoroughly, use a 0.45 μm membrane to filter out impurities before use.
4.3.17. Prepare 100ml blocking buffer (Wester hybridization) 5%(W/V) skimmed milk powder TBST BUffer
(1) Weigh 5g of skimmed milk powder and add it to 100ml of TBST Buffer, stir well to dissolve.
(2) Store at 24°C until use (this sealing solution should be prepared for current use).