36种微生物实验用培养基配制

发布时间:

2022-12-27

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培养基产品购买


36种微生物实验用培养基配制
 
 
 
1.牛肉膏蛋白胨培养基(用于细菌培养)
牛肉膏3g,蛋白胨10g,NaCl 5g,水1000mL,pH7.4~7.6。
 
2.高氏1号培养基(用于放线菌培养)
可溶性淀粉20g,KNO3 1g,NaCl 0.5g,K2HPO4?3H2O 0.5g,MgSO4?7H2O0.5g,FeSO4?7H2O0.01g,水1000mL,pH7.4~7.6。配制时注意:可溶性淀粉要先用冷水调匀后再加入到以上培养基中。
 
3.马丁氏(Martin)培养基(用于从土壤中分离真菌)
K2HPO41g,MgSO4?7H2O0.5g,蛋白胨5g,葡萄糖10g,1/3000孟加拉红水溶液100mL,水900mL,自然pH,121℃湿热灭菌30min。待培养基融化后冷却55~60℃时加入链霉素(链霉素含量为30μg/mL)。
 
4.马铃薯培养基(PDA培养基)(用于霉菌或酵母菌培养)
马铃薯(去皮)200g,蔗糖(或葡萄糖)20g,水1000mL,配制方法如下:
将马铃署去皮,切成约2cm2的小块,放入1500mL的烧杯中煮沸30min,注意用玻棒搅拌以防糊底,然后用双层纱布过滤,取其滤液加糖,再补足至1000mL,自然pH,霉菌用蔗糖,酵母菌用葡萄糖。
 
5.察氏培养基(蔗糖硝酸钠培养基)(用于霉菌培养)
蔗糖30g,NaNO3 2g,K2HPO4 1g,MgSO4?7H2O0.5g,KCl 0.5g,FeSO4?7H2O0.1g,水1000mL,pH7.0~7.2。
 
6.Hayflik培养基(用于支原体培养)
牛心消化液(或浸出液)1000mL,蛋白胨10g,NaCl 5g,琼脂15g,pH7.8~8.0,分装每瓶70mL,121℃湿热灭菌15min,待冷却至80℃左右,每70mL中加入马血清20mL,25%鲜酵母浸出液10mL,15醋酸铊水溶液2.5mL,青霉素G钾盐水溶液(20万单位以上)0.5mL,以上混合后倾注平板。
*注意:醋酸铊是极毒的药品,需特别注意安全操作。
 
7.麦氏(McCLary)培养基(醋酸钠培养基)
葡萄糖0.1g, KCl 0.18g,酵母膏0.25g,醋酸钠0.82g,琼脂l.5g,蒸馏水l00mL。溶解后分装试管,1l5℃湿热灭菌15min。
 
8.葡萄糖蛋白胨水培养基(用于V.P.反应和甲基红试验)
蛋白胨0.5g,葡萄糖0.5g,K2HPO4 0.2g,水100mL,pH7.2,1l5℃湿热灭菌20min。
 
9.蛋白胨水培养基(用于吲哚试验)
蛋白胨10g,NaCl 5g,水1000mL,pH7.2~7.4,121℃湿热灭菌20min。
10.糖发酵培养基(用于细菌糖发酵试验)
蛋白胨0.2g,NaCl 0.5g,K2HPO4 0.02g,水100mL,溴麝香草酚蓝(1%水溶液)0.3mL,糖类lg。分别称取蛋白胨和NaCl溶于热水中,调pH至7.4,再加入溴麝香草酚蓝(先用少量95%乙醇溶解后,再加水配成1%水溶液),加入糖类,分装试管,装量4~5cm高,并倒放入一杜氏小管(管口向下,管内充满培养液)。115℃湿热灭菌20min。灭菌时注意适当延长煮沸时间,尽量把冷空气排尽以使杜氏小管内不残存气泡。常用的糖类,如葡萄糖、蔗糖、甘露糖、麦芽糖、乳糖、半乳糖等(后两种糖的用量常加大为1.5%)。
 
11.RCM培养基(强化梭菌培养基)、(用于厌氧菌培养)
酵母膏3g,牛肉膏l0g,蛋白胨10g,可溶性淀粉lg,葡萄糖5g, 半胱氨酸盐酸盐0.5g,NaCl 3g,NaAc 3g,水l000mL,pH8.5,刃天青3mg/L,l2l℃湿热灭菌30min。
 
12.TYA培养基(用于厌氧菌培养)
葡萄糖40g,牛肉膏2g,酵母膏2g,胰蛋白胨(bacto-typetone)6g,醋酸铵3g,KH2PO4 0.5g,MgSO4?7H2O0.2g,FeSO4?7H2O0.01g,水1000mL, pH6.5,121℃湿热灭菌30min。
 
13.玉米醪培养基(用于厌氧菌培养)
玉米粉65g,自来水1000mL,混匀,煮10min成糊状,自然pH,121℃湿热灭菌30min。
 
14.中性红培养基(用于厌氧菌培养)
葡萄糖40g,胰蛋白胨6g,酵母膏2g,牛肉膏2g,醋酸铵3g,KH2PO4 5g,中性红0.2g,MgSO4?7H2O0.2g,FeSO4?7H2O0.01g, 水1000mL,pH6.2,121℃湿热灭菌30min。
 
15.CaCO3明胶麦芽汁培养基(用于厌氧菌培养)
麦芽汁(6波美)1000mL,水1000mL,CaCO310g,明胶10g,pH6.8,121℃湿热灭菌30min。
 
16.BCG牛乳培养基(用于乳酸发酵)
(A)溶液:
脱脂乳粉100g,水500mL,加入1.6%溴甲酚绿(B.C.G)乙醇溶液1mL,80℃灭菌20min。
(B)溶液:
酵母膏10g,水500mL,琼脂20g, pH6.8,121℃湿热灭菌20min。以无菌操作趁热将(A)、(B)溶液混合均匀后倒平板。
 
17.乳酸菌培养基(用于乳酸发酵)
牛肉膏5g,酵母膏5g,蛋白胨10g,葡萄糖10g,乳糖5g,NaCl 5g,水1000mL,pH6.8,121℃湿热灭菌20min。
 
18.酒精发酵培养基(用于酒精发酵)
蔗糖10g,MgSO4?7H2O 0.5g,NH4NO30.5g,20%豆芽汁2mL,KH2PO4 0.5g,水100mL,自然pH。
 
19.柯索夫培养基(用于钩端螺旋体培养)
优质蛋白胨0.4g, NaCl 0.7g,KCl 0.02g, NaHCO3 0.01g,CaCl 0.02g,KH2PO40.09g,NaH2PO40.48g,蒸馏水500mL,无菌兔血清40mL。
制法:
除兔血清外的其余各成分混合,加热溶解,调pH至7.2,121℃湿热灭菌20min,待冷却后,加入无菌兔血清,制成8%血清溶液,然后分装试管(5~10mL/管),56℃水浴灭活lh后备用。
 
20.豆芽汁培养基
黄豆芽500g,加水1000mL,煮沸lh,过滤后补足水分,121℃湿热灭菌后存放备用,此即为50%的豆芽汁;
用于细菌培养:
10%豆芽汁200mL,葡萄糖(或蔗糖)50g,水800mL,pH7.2~7.4。
用于霉菌或酵母菌培养:
10%豆芽汁200mL,糖50g,水800mL,自然pH。霉菌用蔗糖,酵母菌用葡萄糖。
 
21.LB(Luria—Bertani)培养基(细菌培养,常在分子生物学中应用)
双蒸馏水950mL,胰蛋白胨l0g,NaCl l0g,酵母提取物(bacto- yeast extract)5g,用lmol/L NaOH (约l mL) 调节pH值至7.0,加双蒸馏水至总体积为1L,121℃湿热灭菌30min。
 
含氨苄青霉素LB培养基:
待LB培养基灭菌后冷至50℃左右加入抗生素,至终浓度为80~100mg/L。
22.复红亚硫酸钠培养基(远藤氏培养基)、(用于水体中大肠菌群测定)蛋白胨10g,牛肉浸膏5g,酵母浸膏5g,琼脂20g,乳糖10g,K2HPO40.5g,无水亚硫酸钠5g,5%碱性复红乙醇溶液20mL,蒸馏水1000mL。
 
制作过程:
先将蛋白胨、牛肉浸膏、酵母浸膏和琼脂加入到900mL水中,加热溶解,再加入K2PO4,溶解后补充水至l000mL,调pH至7.2~7.4。随后加入乳糖,混匀溶解后,于115℃湿热灭菌20min。再称取亚硫酸钠至一无菌空试管中,用少许无菌水使其溶解,在水浴中煮沸10min后,立即滴加于20mL 5%碱性复红乙醇溶液中,直至深红色转变为淡粉红色为止。将此混合液全部加入到上述已灭菌的并仍保持融化状态的培养基中,混匀后立即倒平板,待凝固后存放冰箱备用,若颜色由淡红变为深红,则不能再用。
 
23.乳糖蛋白胨半固体培养基(用于水体中大肠菌群测定)
蛋白胨10g,牛肉浸膏5g,酵母膏5g,乳糖10g,琼脂5g,蒸馏水1000mL,pH7.2~7.4,分装试管(l0mL/管),115℃湿热灭菌20min。
 
24.乳糖蛋白胨培养液(用于多管发酵法检测水体中大肠菌群)
蛋白胨10g,牛肉膏3g,乳糖5g,NaCl 5g,蒸馏水l000mL,1.6%溴甲酚紫乙醇溶液lmL。调pH至7.2,分装试管(l0mL/管),并放入倒置杜氏小管,l15℃湿热灭菌20min。
 
25.三倍浓乳糖蛋白胨培养液(用于水体中大肠菌群测定)
将乳糖蛋白胨培养液中各营养成分以扩大3倍加入到l000mL水中,制法同上,分装于放有倒置杜氏小管的试管中,每管5mL,1l5℃湿热灭菌20min。
 
26.伊红美蓝培养基(EMB培养基)(用于水体中大肠菌群测定和细菌转导)
蛋白胨l0g,乳糖10g,K2HPO4 2g,琼脂25g,2%/伊红Y(曙红)水溶液20mL,0.5%美蓝(亚甲蓝)水溶液l3mL,pH7.4。
制作过程:
先将蛋白胨、乳糖、K2HPO4和琼脂混匀,加热溶解后,调pH至7.4,1l5℃湿热灭菌20min,然后加入已分别灭菌的伊红液和美蓝液,充分混匀,防止产生气泡。待培养基冷却到50℃左右倒平皿。如培养基太热会产生过多的凝集水,可在平板凝固后倒置存于冰箱备用。在细菌转导实验中用半乳糖代替乳糖,其余成分不变。
 
27.加倍肉汤培养基(用于细菌转导)
牛肉膏6g,蛋白胨20g,NaCl 10g,水1000mL,pH7.4~7.6。
28.半固体素琼脂(用于细菌转导)琼脂1g,水100mL,121℃湿热灭菌30min。
 
29.豆饼斜面培养基(用于产蛋白酶霉菌菌株筛选)
豆饼100g加水5~6倍,煮出滤汁100mL,汁内加入KH2PO4 0.1%,MgSO4 0.05%,(NH4)2SO4  0.05%,可溶性淀粉2%,pH6,琼脂2%~2.5%。  
 
30.酪素培养基(用于蛋白酶菌株筛选)
分别配制A液和B液。
A液:
称取Na2HPO4?7H2O 1.07g。干酪素4g,加适量蒸馏水,并加热溶解。
B液:
称取 KH2PO4 0.36g,加水溶解。A、B液混合后,加入酪素水解液0.3 mL,加琼脂20g,最后用蒸馏水定容至1000mL。
 
酪素水解液的配制:
1g酪蛋白溶于碱性缓冲液中,加入1%的枯草芽孢杆菌蛋白酶25mL加水至100mL,30℃水解l h。用于配制培养基时,其用量为1000mL,培养基中加入100mL以上水解液。
 
31.细菌基本培养基(用于筛选营养缺陷型)   
Na2HPO4?7H2O 1g,MgSO4?7H2O0.2g,葡萄糖5g,NaCl 5g,K2HPO4lg,水l000mL,pH7.0,1l5℃湿热灭菌30min。
 
32.YEPD培养基(用于酵母原生质体融合)
酵母粉10g,蛋白胨20g,葡萄糖20g,蒸馏水1000mL,pH6.0,115℃湿热灭菌20min。
 
33.YEPD高渗培养基(用于酵母原生质体融合)
在YEPD培养基中加入0.6mol/L的NaCL,3%琼脂。
 
34.YNB基本培养基(用于酵母原生质体融合)
0.67%酵母氮碱基(YNB, 不含氨基酸,Difco),2%葡萄糖,3%琼脂,pH6.2。
另一配方为:
葡萄糖10g(NH4)2SO4 1g,K2HPO40.125g,KHPO4 0.875g,KI 0.0001g,MgSO4?7H2O0.5g,CaCl2?2H2O0.lg,NaCl0.1g,维量元素母液lmL,维生素母液lmL(母液均按常规配制),水1000mL,pH5.8~6.0。
 
35.YNB高渗基本培养基(用于原生质体融合)
在YNB基本培养基中加入0.6mol/LNaCl。
 
36.酚红半固体柱状培养基(用于检查氧与菌生长的关系)
蛋白胨1g,葡萄糖10g, 玉米浆10g,琼脂7g,水1000mL,pH7.2。在调好pH后,加入1.6%酚红溶液数滴,至培养基变为深红色,分装于大试管中,装量约为试管高度的1/2,1l5℃灭菌20min。细菌在此培养基中利用葡萄糖生长产酸,使酚红从红色变成黄色,在不同部位生长的细菌,可使培养基的相应部位颜色改变,但注意培养时间太长,酸可扩散以致不能正确判断结果。
以上各种培养基均可配制成固体或半固体状态,只需改变琼脂用量即可,前者为1.5%~2.0%,后者为0.3%~0.8%。
 
 
 
36 kinds of microorganism were prepared by culture medium
 
 
 
1. Beef paste peptone culture medium (for bacterial culture)
Beef paste 3g, peptone 10g, NaCl 5g, water 1000mL, pH7.4 ~ 7.6.
 
2. Gaoshi 1 medium (for actinomycetes culture)
Soluble starch 20g, kno31g, NaCl 0.5g, K2HPO4?3H2O 0.5g, MgSO4? 7h2o0.5g, FeSO4? 7h2o0.1 g, water 1000mL, pH7.4 ~ 7.6. Note: soluble starch should be mixed with cold water before adding to the above medium.
 
3. Martin medium (used to separate fungi from soil)
K2HPO41g, MgSO4 ? 7 h2o0. 5 g, 5 g peptone, 10 g of glucose, 1/3000 Bangladesh red aqueous solution 100 ml, 900 ml water, natural pH, 121 ℃ hot and humid sterilization 30 min. After being medium melt cooling 55 ~ 60 ℃ with streptomycin (streptomycin content is 30 mu g/mL).
 
4. Potato culture medium (PDA)(for mold or yeast culture)
Potatoes (peeled)200g, sucrose (or glucose)20g, water 1000mL, the preparation method is as follows:
Jingle bell department, peel cut into 2 cm2 small pieces, add 1500 ml beaker boiling in 30 min, stir with a glass rod to prevent the paste, then use double gauze filter, take its filtrate sugar, up to 1000 ml, natural pH, mould with cane sugar, yeast with glucose.
 
5. Chace culture medium (sucrose sodium nitrate culture medium)(for mold culture)
Sucrose 30g, NaNO3 2g, K2HPO4 1g, MgSO4? 7h2o0.5g, KCl 0.5g, FeSO4? 7h2o0.1g, water 1000mL, pH7.0 ~ 7.2.
 
6.Hayflik medium (for mycoplasma culture)
Beef heart digestive juices (or 1000 mL leaching liquid), peptone 10 G, 5 G NaCl, AGAR, 15 G pH7.8 ~ 8.0, packing bottle of 70 mL, 121 ℃ hot and humid sterilization 15 min, stay cool to about 80 ℃, each add horse serum 20 mL to 70 mL, 25% yeast extract 10 mL, 15 thallium acetic acid aqueous solution 2.5 mL, penicillin G potassium solution (0.5 mL, more than 200000 units) blend to pour into tablets.
* note: thallium acetate is a highly toxic drug and special attention should be paid to safe operation.
 
7. McCLary medium (sodium acetate medium)
Glucose 0.1g, KCl 0.18g, yeast extract 0.25g, sodium acetate 0.82g, agar-agar l.5g, distilled water l00mL. Repackaging tube after dissolving, 1 l5 ℃ hot and humid sterilization for 15 min.
 
8. Glucose pepptone water medium (for V.P. reaction and methyl red test)
Peptone 0.5 g and 0.5 g of glucose, K2HPO4 0.5 g, water 100 ml, pH7.2, 20 min 1 l5 ℃ hot and humid sterilization.
 
9. Peptone water culture medium (for indole test)
Peptone 10 g, 5 g, NaCl water, 1000 ml, pH7.2 ~ 7.4, 121 ℃ hot and humid sterilization 20 min.
 
10. Sugar fermentation medium (for bacterial sugar fermentation test)
Peptone 0.2g, NaCl 0.5g, K2HPO4 0.02g, water 100mL, bromothymol blue (1% aqueous solution) 0.3ml, sugar lg. Weigh and peptone and NaCl respectively dissolved in hot water, the pH to 7.4, add bromothymol blue (with a small amount of 95% alcohol solution, then add water to 1% water solution), add sugar, partial shipments in vitro, charge 4 ~ 5 cm high, a duchenne and backwards into the small tube (tube, down tube filled with culture medium). 115 ℃ hot and humid sterilization 20 min. When sterilizing, pay attention to extend the boiling time properly and exhaust the cold air to keep the bubbles in the tubule. Common sugars, such as glucose, sucrose, mannose, maltose, lactose, galactose, etc.
 
11.RCM culture medium (enhanced clostridium culture medium), (used for anaerobic culture)
Yeast extract, 3 g beef paste l0g, peptone 10 g, soluble starch lg, glucose, 5 g, 0.5 g cysteine hydrochloride, NaCl, 3 g NaAc 3 g, water l000mL pH8.5, resazurin 3 mg/L, l2l ℃ damp heat sterilization for 30 min.
 
12.TYA culture medium (for anaerobic bacteria culture)
Glucose, 40 g beef paste 2 g, 2 g, yeast extract tryptone (bacto - typetone) 6 g, 3 g, ammonium acetate KH2PO4 0.5 g, MgSO4, 7 h2o0. 2 g, FeSO4, 7 h2o0. 01 g, water 1000 ml, pH6.5, 121 ℃ hot and humid sterilization 30 min.
 
13. Culture medium of corn mash (for anaerobic bacteria culture)
Corn flour, 65 g water, 1000 ml, blending, cook for 10 min into paste, natural pH, 121 ℃ hot and humid sterilization 30 min.
 
14. Neutral red medium (for anaerobic culture)
Glucose, 40 g, tryptone 6 g, 2 g yeast extract, beef extract 2 g, 3 g, ammonium acetate KH2PO4 5 g, 0.2 g, neutral red MgSO4 ? 7 h2o0. 2 g, FeSO4, 7 h2o0. 01 g, water 1000 ml, pH6.2, 121 ℃ hot and humid sterilization 30 min.
 
15.CaCO3 gelatin malt juice culture medium (for anaerobic bacteria culture)
Wort (6 wave beauty) 1000 ml, 1000 ml water, CaCO310g, gelatin, 10 g pH6.8, 121 ℃ hot and humid sterilization 30 min.
 
16.BCG milk culture medium (for lactic acid fermentation)
(A) solution:
Dried skim milk, 100 g, 500 ml water, add 1.6% bromocresol green (B.C.G) ethanol solution 1 ml, 80 ℃ sterilization 20 min.
(B) solution:
Yeast extract 10 g, water 500 ml, AGAR, 20 g pH6.8, 121 ℃ hot and humid sterilization 20 min. Mix (A) and (B) solutions thoroughly and pour over the plate.
 
17. Lactobacillus culture medium (for lactic acid fermentation)
Beef extract 5 g, 5 g yeast extract, peptone 10 g, 10 g of glucose, lactose, 5 g, 5 g, NaCl water 1000 ml, pH6.8, 121 ℃ hot and humid sterilization 20 min.
 
18. Alcoholic fermentation medium (for alcoholic fermentation)
Sucrose 10g,MgSO4?7H2O 0.5g, nh4no30.5g,20% bean sprout juice 2mL, KH2PO4 0.5g, water 100mL, natural pH.
 
19. Kosoff culture medium (for leptospira culture)
High-quality peptone 0.4g, NaCl 0.7g,KCl 0.02g, NaHCO3 0.01g, CaCl 0.02g, kh2po40.09g, nah2po40.48g, distilled water 500mL, aseptic rabbit serum 40mL.
Method:
Besides rabbit serum of the rest of the ingredients, cooking, adjust pH value to 7.2, 121 ℃ hot and humid sterilization 20 min, after being cooled, join the sterile rabbit serum, made from 8% serum solution, and in vitro (5 ~ 10 ml/pipe), 56 ℃ water bath inactivated lh backup.
 
20. Bean sprout juice culture medium
Yellow bean sprouts 500 g, 1000 ml of water, boil lh, supply water after filtration, 121 ℃ after damp heat sterilization for backup, this is for 50% of the bean sprout juice;
For bacterial culture:
10% bean sprouts juice 200mL, glucose (or sucrose)50g, water 800mL, pH7.2 ~ 7.4.
For mold or yeast culture:
10% bean sprouts juice 200mL, 50g sugar, 800mL water, natural pH. Molds use sucrose, yeast USES glucose.
 
21.LB(luria-bertani) medium (bacterial culture, often used in molecular biology)
Double distilled water, 950 mL, tryptone l0g, NaCl l0g, yeast extract (bacto - yeast extract), 5 g with lmol/L NaOH (about L mL) to adjust pH value to 7.0, double distilled water to the total volume of 1 L, 121 ℃ hot and humid sterilization 30 min.
 
LB medium containing ampicillin:
Stay LB medium after sterilization cold to 50 ℃ or so to join the antibiotics, and concentration of 80 ~ 100 mg/L.
22. The complex red (endo's medium), sodium sulfite medium (applied to determination of coliform bacteria in water) peptone 10 g, 5 g beef extract, yeast extract, 5 g, AGAR 20 g, 10 g, lactose K2HPO40. 5 g, 5 g anhydrous sodium sulfite, 5% ethanol solution of alkaline complex red 20 ml, distilled water, 1000 ml.
 
Production process:
Peptone, beef extract, yeast extract and AGAR AGAR were first added to 900mL water, which was heated and dissolved, followed by K2PO4. After dissolution, water was added to l000mL and pH was adjusted to 7.2 ~ 7.4. Add the lactose, blending dissolves, 20 min at 115 ℃ hot and humid sterilization. Said again take sodium sulfite to a sterile empty tube, with a few sterile water to dissolve it, after the water boil for 10 min, immediately drop founded more than 20 ml 5% ethanol solution of alkaline complex red, until turned into deep red to pale pink. And sterilized to add all the mixture to the above and still keep the melting state of medium, immediately after each tablet, after being frozen storage refrigerator spare, if the color changed from pink to scarlet, cannot reoccupy.
 
23. Lactose peptone semi-solid medium (for the determination of coliform flora in water)
Peptone 10 g, 5 g beef extract, yeast extract, 5 g lactose 10 g, 5 g AGAR, distilled water, 1000 ml, pH7.2 ~ 7.4, repackaging tube (l0mL/tube), 115 ℃ hot and humid sterilization 20 min.
 
24. Lactose peptone culture solution (for the detection of coliform bacteria in water by multi-tube fermentation)
Peptone 10g, beef paste 3g, lactose 5g, NaCl 5g, distilled water l000mL, 1.6% bromocresol violet ethanol solution lmL. Adjust pH to 7.2, repackaging tube (l0mL/tube), and into the inversion duchenne tubular, l15 ℃ hot and humid sterilization 20 min.
 
25. Triplose-concentrated lactose peptone culture solution (for the determination of coliform flora in water)
Will lactose peptone medium, the nourishment composition to expand 3 times, joining l000mL water method as above, packing in the test tube with our horse duchenne tubular, each tube 5 ml, 1 l5 ℃ hot and humid sterilization 20 min.
 
26. Erythrocyte methylene blue culture medium (EMB culture medium)(for the determination of coliforms and bacterial transduction in water)
Peptone l0g, lactose 10g, K2HPO4 2g, agar-agar 25g, 2%/ eosin Y(shu hong) aqueous solution 20mL,0.5% methylene blue (methylene blue) aqueous solution l3mL, pH7.4.
Production process:
Lactose peptone, first, K2HPO4 and AGAR blending, heating dissolves, pH 7.4, 1 l5 ℃ hot and humid sterilization 20 min, and then joined the sterilization of red liquid and blue liquid respectively, thoroughly inc