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1、实验室常用贮存液的配制参数（Preparation parameters of laboratory common storage liquid）Preparation parameters of laboratory common storage liquidview the data uploaded by this user in the data centerpersonal data give him a message post collectionreply citation maintenance-1.30% acrylamide solutionpreparation met
2、hod 29g acrylamide and 1g, N, N - bis - dimethyl acrylamide were dissolved in the water of 60ml in total volume. Heat up to 37 DEG C, dissolve, add water to the final volume of 100ml. Nalgene filter (0.45 m pore diameter) was used to remove bacteria, and the pH value of the solution should be no mor
3、e than 7. Keep the brown bottle at room temperature.note acrylamide has strong neurotoxicity and can be absorbed through the skin, and its effects are cumulative. Gloves and masks should be worn when weighing acrylamide and methylene acrylamide. Polyacrylamide may be considered non-toxic, but it sho
4、uld be handled with care because it may contain a small amount of non polymeric material. Some of the low price of acrylamide and bisacrylamide usually contain some metal ions in the storage of acrylamide single bed mixed resin added about 0.2 volume of fluid (MB-1Mallinckrodt), stirring overnight,
5、and then use the Whatman filter to purify the No. 1. During storage, acrylamide and acrylamide are slowly converted to propylene and double acrylic acid.2.40% acrylamidepreparation method 380g acrylamide (DNA sequencing stage) and 20g, N, N - bis - dimethyl acrylamide were dissolved in distilled wat
6、er with a total volume of 600ml. Continue to prepare 30% acrylamide solution according to the above process, but after heating dissolved, should be distilled water to make up to the final volume of 1L.note see the above formulation of 30% acrylamide instructions, 40% acrylamide solution for DNA sequ
7、encing.3. actinomycin D solutionmethod 20mg actinomycin D was dissolved in 4ml 100% ethanol, diluted at 1:10, and 100% ethanol was used as blank control to read OD440 value. Actinomycin D (molecular weight 1255) Moore pure digestibility coefficient in aqueous solution was 21900, the actinomycin D so
8、lution absorbance at 440nm was 0.182 and 1mg/ml, the storage solution of actinomycin D should be placed with the foil in the package tube, stored in the -20 C.note actinomycin D is teratogens and carcinogens, must wear gloves and operating under the hood of the preparation solution, but not in the o
9、pen in the experiment on the desktop, beware of inhalation powder or let it come into contact with the eyes or skin.Actinomycin D products provided by pharmaceutical companies for therapeutic use often contain additives such as sugar or salt. By measuring the concentration of actinomycin D in the st
10、orage fluid at the light of the 440nm wave strengths, such products can be used to inhibit their own guiding effects.4.0.1mol/L adenosine three phosphate (ATP) solutionmethod of preparation dissolve 60mg, ATP in 0.8ml water, use 0.1mol/L NaOH to adjust pH value to 7, use distilled water constant vol
11、ume 1ml, pack into small parts, keep at -70 degrees centigrade5.10mol/L acetic acid solutionpreparation method dissolve 770g acetic acid in 800ml water, then add water to 1L, then filter and sterilize.6.10% ammonium persulfate solutionmethod 1g ammonium persulfate was dissolved in an aqueous solutio
12、n of 10ml at last, and the solution could be stored for 4 weeks at a temperature of several weeks.7.BCIP solutionmethod 0.5g 5-, bromine, -4-, chloride, -3-, indole, phosphoric acid, two sodium salt (BCIP) were dissolved in two methyl formamide of 10ml 100%, stored at 4 DEG C8.2 * BES buffer salt so
13、lutionMethods prepared with distilled water dissolved 1.07g salt solution BESN total volume of 90ml, N- bis (2- hydroxyethyl) -2- Aminoethylsulfonic acid, 1.6g NaCl and 0.027g Na2HPO4, room temperature regulation of the solution with HCl pH value to 6.96, then add distilled water volume to 100ml, wi
14、th 0.22 M filter sterilization, packing into small portions, stored in the -20 C.9.1mol/L CaCl2 solutionmethod of preparation dissolve 54G, CaCl2, 6H2O in 200ml distilled water, filter with 0.22 mu m filter, strain it into small parts of 10ml and store at -20 DEG C.note preparing competent cells, re
15、move a small portion of thawed and diluted with distilled water to 100ml, using the Nalgene filter (0.45 m diameter) filtration, then quenched to 0 DEG C.10.2.5mol/L CaCl2 solutionmethod of preparation dissolve 13.5g, CaCl2, 6H2O in 20ml distilled water, filter with 0.22 mu m filter, strain it into
16、small parts of 1ml and store at -20 DEG C.11.1mol/L, two, DTT solutionmethod 20ml 0.01mol/L sodium acetate solution (pH5.2) was used to dissolve 3.09g and DTT. After filtration, the bacteria were separated into small parts of 1ml and stored at -20 DEG C.note DTT or solutions containing DTT should no
17、t be subjected to high-pressure treatment.12. deoxy nucleoside three phosphoric acid (dNTP) solutionmethod the preparation of each kind of dNTP dissolved in water to the concentration is about 100mmol/L, with a pipette from 0.05mol/l Tris base respectively to control each dNTP pH value of the soluti
18、on 7 (pH test), each dNTP solution after neutralization to take one each for proper dilution in the the wavelength of reading optical density to calculate the actual concentration of each dNTP, and then diluted into the final concentration of 50mmol/L dNTP, and packaged into small portions and store
19、d in the -70 C.13.0.5mol/l, EDTA (pH8.0) solutionMethods with adding two 186.1g water two 800ml EDTA sodium in water (EDTA-Na? 2H2O), vigorous stirring on a magnetic stirrer, the solution is adjusted by NaOH pH to 8 (about 20g NaOH particles) and then set the volume at 1L, after filling high-pressur
20、e sterilization standby.note EDTA two sodium salt needs to be added to NaOH to raise the pH value of the solution to nearly 8, so that it can be completely dissolved.14. ethidium bromide (10mg/ml solution)method with 1g preparation of ethidium bromide in 100ml water, stirring for several hours to en
21、sure that it is completely dissolved, and then transferred to the container with aluminum foil wrapped or brown bottle, stored at room temperature.note caution: ethidium bromide is a strong mutagen and has moderate toxicity, wear gloves using a solution containing this dye, to wear a mask when weigh
22、ing dyes.15.2 * HEPES buffer salt solutionmethod of preparation dissolve 1.6g, NaCl, 0.074g, KCl, 0.027g, Na2PO4, 2H2O, 0.2g, dextran and 1gHEPES with a total amount of 90ml of distilled water, adjust the pH value to 7.05 with 0.5mol/l NaOH, and then use distilled water to 100ml. The filter is steri
23、lized by 0.22 M filter and divided into small parts of 5ml, stored at -20 DEG C.16.IPTG solutionmethod IPTG for preparation of isopropyl thio - -D- beta galactosidase (molecular weight 238.3), 2G IPTG dissolved in 8ml of distilled water, distilled water volume to 10ml, with 0.22 M filter sterilizati
24、on, packing into 1ml pieces stored in -20 C.17.1mol/L magnesium acetate solutionmethod of preparation dissolve 214.46g, four mg of water in 800ml water, use water to 1L, filter and sterilize.18.1mol/L MgCl2 solutionmethod of preparation dissolve 203.4g, MgCl2, 6H2O in water of 800ml, water content t
25、o 1L, sub packed into small parts and sterilized by high pressure.note MgCl2 is deliquescent, should choose the vial (such as 100g) reagent, the opening of the new bottle after the not long-term storage.19. BME solutionpreparation method generally obtained 14.4mol/L solution, should be installed in
26、brown bottle, stored at 4 degrees centigrade.note BME or solutions containing BME should not be treated with high pressure.20.NBT solutionpreparation method 0.5g tetrazolium chloride four tetrazolium was dissolved in two methyl formamide of 10ml 70%, stored at 4 DEG C.Related reagents: chemical reag
27、ents, standard substances, biochemical reagents2006-10-16 20:00:001 floor: RE: share laboratory common storage fluid preparation parametersview the data uploaded by this user in the data center to give him a message of personal data collection postreply citation maintenance-21. phenol / chloroform s
28、olutionmethod the preparation of phenol and chloroform after mixing with 0.1mol/L Tris? HCl (pH7.6) from several times to balance the mixture, the brown glass bottle, covered with equal volume of 0.01mol/l Tris? HCl (pH7.6) liquid layer, preserved at the temperature of 4.note phenol is very corrosiv
29、e and can cause serious burns. Gloves and goggles should be worn while wearing protective clothing. All operations shall be carried out in a chemical fume hood. Wash skin with a large amount of water and wash with soap and water, and avoid using alcohol.22.10mmol/L methyl sulfonyl fluoride (PMSF) so
30、lutionpreparation method dissolve PMSF into 1.74mg/ml (10mmol/L) with isopropanol, then pack it into small parts and store at -20 DEG C. If necessary, a storage solution (100mmol/L) with a concentration of up to 17.4mg/ml can be prepared.note PMSF seriously damages the mucous membranes of the respir
31、atory tract, eyes and skin, and is fatal after inhalation, swallowing, or absorption through the skin. Once the eye or skin is exposed to PMSF, rinse immediately with plenty of water. Any clothing contaminated by PMSF shall be discarded. PMSF is unstable in aqueous solution. Should be used prior to
32、use, from storage liquid, into the pyrolysis buffer. The loss rate of PMSF activity in aqueous solution increases with the increase of pH value, and the inactivation rate at 25 DEG C is higher than 4. When the pH value is 8, the half-life of 20 mmol/l PMSF aqueous solution is about 85min, indicating
33、 that the PMSF solution is adjusted to alkaline (pH8.6) and can be safely discarded for several hours at room temperature.23. phosphate buffer solution (PBS) solutionmethod in the preparation of 800ml distilled water dissolved 8g NaCl, 0.2g KCl, 1.44g Na2HPO4 and 0.24g KH2PO4, the solution is adjust
34、ed by HCl pH to 7.4 water volume to 1L, in 15lbf/in2 (1034 x 105Pa) under high pressure steam sterilization 20min. Keep it at room temperature.24.1mol/L potassium acetate (pH7.5) solutionpreparation method dissolve 9.82g potassium acetate in 90ml pure water, adjust the pH value to 7.5 with 2mol/L ac
35、etic acid, add pure water to 1L, and keep at -20 DEG C.25. potassium acetate solution (used for alkaline cracking)method 11.5ml, glacial acetic acid and 28.5ml water were added to 60ml 5mol/L potassium acetate solution, i.e., the solution with potassium concentration of 3mol/L and acetic acid concen
36、tration of 5mol/L.26.3mol/L sodium acetate (pH5.2 and pH7.0) solutionmethod dissolve 408.1g trihydrate sodium acetate in 80ml water, adjust pH value to 5.2 by glacial acetic acid, or adjust pH value to 7 with dilute acetic acid, add water to 1L, and sterilize after packing.27.5mol/L NaCl solutionmet
37、hod dissolve 292.2g NaCl in 800ml water and add water to 1L. After packing, high pressure sterilization.28.10%, twelve alkyl sodium sulfate (SDS) solutionpreparation method dissolve 100g electrophoresis grade SDS in 900ml water, heat up to 68 DEG C, dissolve in solution, add a few drops of concentra
38、ted hydrochloric acid, adjust the pH value of the solution to 7.2, add water to 1L, then equip.note the fine grain of SDS is easy to diffuse, so it is necessary to wear a face mask when weighing. After weighing, the SDS and 10%SDS solution remaining in the weighing work area and the balance are elim
39、inated.29.20 x SSC solutionpreparation method dissolve 175.3g, NaCl and 88.2g sodium citrate in 800ml water, add a few drops of 10mol/l NaOH solution, adjust pH value to 7, add water constant volume to 1L, after packing, high pressure sterilization.30.20 x SSPE solutionmethod of preparation dissolve
40、 17.5g, NaCl, 27.6g, NaH2PO4, H2O and 7.4g EDTA in 800ml water, adjust pH value to 7.4 by using NaOH solution (about 6.5ml 10ml/L NaOH), add water to 1L, and sterilize after partial loading.31.100%, three chloroacetic acid solutionmethod 227ml water is added to the bottle containing 500g TCA, and th
41、e solution formed contains 100% (M/V) TCA.32.1mol/L Tris solutionmethod of preparation dissolve 121.91g Tris base in 800ml water, add concentrated HCl, adjust pH value to the required value. Should be cold until the room temperature can be adjusted at the end of the pH value, add water constant volu
42、me to 1L, after packing, high pressure sterilization.note yellow solution such as 1mol/L, should be discarded and a better quality Tris.Although many types of electrodes can not accurately measure the pH value of Tris solutions, they can still be purchased from most manufacturers. The pH value of th
43、e Tris solution varies with temperature, and the temperature is increased by 1 DEG C, and the pH value is reduced by about 0.03 units. For example, the pH values at 5, 25, and 37 DEG 0.05mol/L are 9.5, 8.9, and 8.6, respectively.33.Tris buffered saline solution (TBS) (25mmol/l, Tris)method in the pr
44、eparation of 800ml distilled water dissolved 8g NaCl, 0.2g KCl and 3G Tris base, adding 0.015g and HCl to phenol pH value to 7.4, volume to 1L with distilled water, after filling in 151bf/in2 (1.034 x 105Pa) under high pressure steam sterilization 20min at room temperature.34.X-gal solutionpreparati
45、on method X-gal is 5-, bromine, -4-, chloride, -3-, indole - beta, -D, galactosidase. Storage fluid of 20mg/ml prepared by dissolving X-gal with two base formamide. Stored in a glass tube or polypropylene tube, a test tube containing X-gal solution shall be wrapped with aluminum foil to prevent damage due to exposure to light and stored at -20 degrees centigrade. X-gal solution needs no filtration and sterilization.