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      請問:實驗室常用培養基配方有哪些?

      發布時間:

      2022-12-24

      作者:

      中國培養基做的比較大的公司


      請問:實驗室常用培養基配方有哪些?
       
       
      2xYT培養基
        組份濃度 1.6%(W/V)Tryptone,1%(W/V)Yeast extract,0.5%(W/V)NaCl
        配制量 1L
        配制方法
      1.稱量16g Tryptone,10g Yeast extract,5g NaCl置于1L燒杯中。
      2.加入約800ml的去離子水,充分攪拌溶解。
      3.滴加5N NaOH,調節pH值至7.0。
      4.加去離子水將培養基定容至1L。
      5.高溫高壓滅菌后,4℃保存。
       
       bx broth培養基
        組份濃度 2%(W/V) Tryptone,0.5%(W/V) Yeast extract,0.5%(V/V) MgSO4•7H2O。
        配制量 1L
        配制方法
      1.稱量20g Tryptone,5g Yeast extract,5g MgSO4•7H2O置于1L燒杯中。
      2.加入約800ml的去離子水,充分攪拌溶解。
      3.滴加5N NaOH,調節pH值至7.5。
      4.加去離子水將培養基定容至1L。
      5.高溫高壓滅菌后,4℃保存。
       
       NZCYM培養基
        組份濃度 0.5%(W/V) Yeast extract,0.1%(W/V) Casamino Acid,1%(W/V) NZ胺,0.5%(W/V) NaCl,0.2%(W/V) MgSO4•7H2O。
        配制量 1L
        配制方法
      1.稱量5g Yeast extract,1g Casamino Acid,10g NZ胺,5g NaCl,2g MgSO4•7H2O置于1L燒杯中。
      2.加入約800ml的去離子水,充分攪拌溶解。
      3.滴加5N NaOH,調節pH值至7.0。
      4.加去離子水將培養基定容至1L。
      5.高溫高壓滅菌后,4℃保存。
       
       NZYM培養基
        組份濃度 0.5%(W/V) Yeast extract,1%(W/V) NZ胺,0.5%(W/V) NaCl,0.2%(W/V) MgSO4•7H2O。
        配制量 1L
        配制方法
      1.稱量5g Yeast extract,10g NZ胺,5g NaCl,2g MgSO4•7H2O置于1L燒杯中。
      2.加入約800ml的去離子水,充分攪拌溶解。
      3.滴加5N NaOH,調節pH值至7.0。
      4.加去離子水將培養基定容至1L。
      5.高溫高壓滅菌后,4℃保存。
       
       NZM培養基
        組份濃度 1%(W/V)NZ胺,0.5%(W/V) NaCl,0.2%(W/V) MgSO4•7H2O。
        配制量 1L
        配制方法
      1.稱量10g NZ胺,5g NaCl,2g MgSO4•7H2O置于1L燒杯中。
      2.加入約800ml的去離子水,充分攪拌溶解。
      3.滴加5N NaOH,調節pH值至7.0。
      4.加去離子水將培養基定容至1L。
      5.高溫高壓滅菌后,4℃保存。
       
       一般固體培養基
        配制方法
      1.按照液體培養基配方準備好液體培養基,在高溫高壓滅菌前,加入15g/L的Agar(瓊脂:鋪制平板用)或7g/L的Agar(瓊脂:鋪制頂層瓊脂用)或15g/L的Agarose(瓊脂糖:鋪制平板用)或7g/L的Agarose(瓊脂糖:鋪制頂層瓊脂用)。
      2.高溫高壓滅菌后,戴上手套取出培養基,搖動容器使瓊脂或瓊脂糖充分混勻。
      3.待培養基冷卻至50-60℃,加入熱不穩定物質(如,抗生素等),搖動混勻。
      4.鋪板(30-35ml培養基/90mm培養皿)。
       
       LB/Amp/X-Gal/IPTG平板培養基
        組份濃度 1%(W/V) Trytone,0.5%(W/V) Yeast extract, 1%(W/V) NaCl,0.1mg/ml Ampicillin,0.024mg/ml IPTG,0.04mg/ml X-Gal,1.5%(W/V) Agar。
        配制量 1L
        配制方法
      1.稱取10g Tryptone,5g Yeast extract,10g NaCl置于1L燒杯中。
      2.加入約800ml的去離子水,充分攪拌溶解。
      3.滴加5N NaOH,調節pH值至7.0。
      4.加去離子水將培養基定容至1L后,加入15g Agar。
      5.高溫高壓滅菌后,冷卻至60℃左右。
      6.加入1ml Ampicillin(100mg/ml)、1ml IPTG(24 mg/ml)、2ml X-Gal (20mg/ml)后均勻混勻。
      7.鋪板(30-50ml/90mm培養皿)。
      8.4℃避光保存。
       
       TB/Amp/X-Gal/IPTG平板培養基
        組份濃度 1.2%(W/V) Tryptone,2.4%(W/V) Yeast extract,0.4%(W/V) Glycerol,17m M KH2PO4,72mM K2HPO4, 0.1 mg/ml Ampicillin,0.024mg/ml IPTG,0.04mg/ml X-Gal,1.5%(W/V) Agar。
        配制量 1L
        配制方法
      1.配制磷酸鹽緩沖液(0.17 M KH2PO4 , 0.72M K2HPO4) 100ml。
      溶解2.31g KH2PO4 和12.54g K2HPO4 于90ml的去離子水中,攪拌溶解后,加去離子水定容至100ml,高溫高壓滅菌。
      2.稱量12g Tryptone,24g Yeast extract,4ml Glycerol置于1L燒杯中。
      3.加入約800ml的去離子水,充分攪拌溶解并定容至1L后,加入15g Agar,高壓高溫滅菌。
      4.待溶液冷卻至60℃以下時,加入100ml的上述滅菌磷酸鹽緩沖液和1ml的Ampicillin (100mg/ml)、1ml IPTG (24mg/ml)和2ml X-Gal (20mg/ml)。
      5.鋪板(30-35ml培養基/90mm培養皿)。
      6.均勻混合后4℃避光保存。
       
      Ampicillin (100mg/ml)
        組份濃度 100mg/ml Ampicillin
        配制量 50ml
        配制方法
      1.稱量5g Ampicillin置于50ml離心管中。
      2.加入40ml滅菌水,充分混合溶解后,定容至50ml。
      3.用0.22um 濾器過濾滅菌。
      4.小份分裝(1ml/份)后,-20℃保存。
       
       IPTG (24mg/ml)
        組份濃度 24mg/ml IPTG
        配制量 50ml
        配制方法
      1.稱量1g X-Gal置于50ml 離心管中。
      2.加入40ml DMF(二甲基甲酰胺),充分混合溶解后,定容至50ml。
      3. 小份分裝(1ml/份)后,-20℃避光保存。
       
       LB培養基
        組份濃度 1%(W/V) Tryptone,0.5%(W/V) Yeast extract,1%(W/V) NaCl
        配制量 1L
        配制方法
      1.稱量10g Tryptone,5g Yeast extract,10g NaCl。
      2.加入800ml的去離子水,充分攪拌溶解。
      3.滴加5N NaOH (約0.2ml),調節pH值至7.0。
      4.加去離子水將培養基定容至1L。
      5.高溫高壓滅菌后,4℃保存。
       
       LB/Amp培養基
        組份濃度 1%(W/V) Tryptone,0.5%(W/V) Yeast extract,1%(W/V) NaCl,0.1 mg/ml Ampicillin
        配制量 1L
        配制方法
      1.稱量10g Tryptone,5g Yeast extract,10g NaCl。
      2.加入800ml的去離子水,充分攪拌溶解。
      3.滴加5N NaOH(約0.2ml),調節pH值至7.0。
      4.加去離子水將培養基定容至1L。
      5.高溫高壓滅菌后,冷卻至室溫。
      6.加入1ml Ampicillin(100mg/ml)均勻混合后4℃保存。
       
       TB培養基
        組份濃度 1.2%(W/V) Tryptone,2.4%(W/V) Yeast extract,0.4%(V/V) Glycerol,17mM KH2PO4,72mM K2HPO4
        配制量 1L
        配制方法
      1.配制磷酸鹽緩沖液(0.17 M KH2PO4 , 0.72 M K2HPO4) 100ml。
      溶解2.31g KH2PO4 和12.54g K2HPO4 于90ml的去離子水中,攪拌溶解后,加去離子水定容至100ml,高溫高壓滅菌。
      2.稱量12g Tryptone,24g Yeast extract,4ml Glycerol置于1L燒杯中。
      3.加入約800ml的去離子水,充分攪拌溶解并定容至1L 后,高壓高溫滅菌。
      4.待溶液冷卻至60℃以下時,加入100ml的上述滅菌磷酸鹽緩沖液。
      5.4℃保存。
       
       TB/Amp培養基
        組份濃度 1.2%(W/V) Tryptone,2.4%(W/V) Yeast extract,0.4%(W/V) Glycerol,17 mM KH2PO4,72mM K2HPO4 ,0.1 mg/ml Ampicillin
        配制量 1L
        配制方法
      1.配制磷酸鹽緩沖液(0.17 M KH2PO4 , 0.72M K2HPO4) 100ml。
      溶解2.31g KH2PO4 和12.54g K2HPO4 于90ml的去離子水中,攪拌溶解后,加去離子水定容至100ml,高溫高壓滅菌。
      2.稱量12g Tryptone,24g Yeast extract,4ml Glycerol置于1L燒杯中。
      3.加入約800ml的去離子水,充分攪拌溶解并定容至1L后,高壓高溫滅菌。
      4.待溶液冷卻至60℃以下時,加入100ml的上述滅菌磷酸鹽緩沖液和1ml的Ampicillin(100mg/ml)。
      5.均勻混合后4℃保存。
       
       SOB培養基
        組份濃度 2%(W/V) Tryptone,0.5%(W/V) Yeast extract,0.05%(W/V) NaCl,2.5mM KCl,10mM MgCl2
        配制量 1L
        配制方法
      1.配制250mM KCl溶液。
      在90ml的去離子水中溶解1.86g KCl后,定容至100ml。
      2.配制2M MgCl2 溶液。
      在90ml的去離子水中溶解19g MgCl2后,定容至100ml,高溫高壓滅菌。
      3.稱量20g Tryptone,5g Yeast extract,0.5g NaCl置于1L燒杯中。
      4.加入約800ml的去離子水,充分攪拌溶解。
      5.量取10ml 250mM KCl溶液,加入到燒杯中。
      6.滴加5N NaOH溶液(約0.2ml),調節pH值至7.0。
      7.加入去離子水將培養基定容至1L。
      8.高溫高壓滅菌后4℃保存。
      9.使用前加入5ml滅菌的2M MgCl2。
       
       SOC培養基
        組份濃度 2%(W/V) Tryptone,0.5%(W/V) Yeast extract,0.05%(W/V) NaCl,2.5mM KCl,10mM MgCl2,20mM 葡萄糖
        配制量 100ml
        配制方法
      1.配制1M葡萄糖溶液。
      將18g 葡萄糖溶于90ml去離子水中,充分溶解后定容至100ml,用0.22um濾器過濾除菌。
      2.向100ml SOB培養基中加入除菌的1M葡萄糖溶液2ml,均勻混合后4℃保存。
       
       

      Laboratory commonly used medium formula
       
       
      2 xyt medium
      The concentration of the components was 1.6%(W/V)Tryptone, 1%(W/V) calcium extract, 0.5%(W/V)NaCl
      Dispensing quantity 1 l
      Preparation methods
      1. Weigh 16g Tryptone, 10g torus extract, 5g NaCl in a 1L beaker.
      2. Add about 800ml of deionized water and stir well to dissolve.
      3. Add 5N NaOH drops to adjust pH to 7.0.
      4. Add deionized water to set the medium capacity to 1L.
      5. After the high temperature and high pressure sterilization, 4 ℃.
       
      Bx broth broth medium
      The concentration of the components is 2%(W/V) Tryptone, 0.5%(W/V) calcium extract, 0.5%(V/V) MgSO4•7H2O.
      Dispensing quantity 1 l
      Preparation methods
      1. Weigh 20g Tryptone, 5g torajin, 5g MgSO4•7H2O and place them in 1L beaker.
      2. Add about 800ml of deionized water and stir well to dissolve.
      3. Add 5N NaOH drops to adjust pH value to 7.5.
      4. Add deionized water to set the medium capacity to 1L.
      5. After the high temperature and high pressure sterilization, 4 ℃.
       
      NZCYM medium
      The concentration of components is 0.5%(W/V), 0.1%(W/V) Casamino Acid, 1%(W/V) NZ amine, 0.5%(W/V) NaCl, 0.2%(W/V) MgSO4•7H2O.
      Dispensing quantity 1 l
      Preparation methods
      1. 5g Osaka extract, 1g Casamino Acid, 10g NZ amine, 5g NaCl, and 2g MgSO4•7H2O were weighed in 1L beaker.
      2. Add about 800ml of deionized water and stir well to dissolve.
      3. Add 5N NaOH drops to adjust pH to 7.0.
      4. Add deionized water to set the medium capacity to 1L.
      5. After the high temperature and high pressure sterilization, 4 ℃.
       
      NZYM medium
      The concentration of components is 0.5%(W/V), 1%(W/V) NZ amine, 0.5%(W/V) NaCl, 0.2%(W/V) MgSO4•7H2O.
      Dispensing quantity 1 l
      Preparation methods
      1. 5g Osaka extract, 10g NZ amine, 5g NaCl, and 2g MgSO4•7H2O were weighed and placed in a 1L beaker.
      2. Add about 800ml of deionized water and stir well to dissolve.
      3. Add 5N NaOH drops to adjust pH to 7.0.
      4. Add deionized water to set the medium capacity to 1L.
      5. After the high temperature and high pressure sterilization, 4 ℃.
       
      NZM medium
      The concentration of components was 1%(W/V)NZ amine, 0.5%(W/V) NaCl, 0.2%(W/V) MgSO4•7H2O.
      Dispensing quantity 1 l
      Preparation methods
      1. Weigh 10g NZ amine, 5g NaCl and 2g MgSO4•7H2O and place them in 1L beaker.
      2. Add about 800ml of deionized water and stir well to dissolve.
      3. Add 5N NaOH drops to adjust pH to 7.0.
      4. Add deionized water to set the medium capacity to 1L.
      5. After the high temperature and high pressure sterilization, 4 ℃.
       
      General solid medium
      Preparation methods
      1. According to the fluid medium formula prepared liquid medium, in front of the high temperature and high pressure sterilization, join 15 g/L of Agar (Agar: spread sheet use) or 7 g/L of Agar (top Agar Agar: spread system) or 15 g/L of Agarose (Agarose: spread sheet use) or 7 g/L of Agarose (top Agar with Agarose: spread system).
      2. After sterilization at high temperature and high pressure, put on gloves and take out the culture medium. Shake the container to fully mix AGAR or agarose.
      3. Stay cool medium to 50 to 60 ℃, add hot unstable substances (e.g., antibiotics, etc.), shaking the blender.
      4. Planking (30-35ml medium /90mm dish).
       
      LB/Amp/ x-gal /IPTG flat medium
      The concentration of components is 1%(W/V) Trytone, 0.5%(W/V) extract, 1%(W/V) NaCl, 0.1mg/ml Ampicillin, 0.024mg/ml IPTG, 0.04mg/ml x-gal, 1.5%(W/V) Agar.
      Dispensing quantity 1 l
      Preparation methods
      1. It is said that 10g Tryptone, 5g Osaka extract and 10g NaCl were placed in a 1L beaker.
      2. Add about 800ml of deionized water and stir well to dissolve.
      3. Add 5N NaOH drops to adjust pH to 7.0.
      4. Add 15g Agar after removing the ionized water and setting the medium to 1L.
      5. After the high temperature and high pressure sterilization, cooling and 60 ℃ or so.
      6. Add 1ml Ampicillin(100mg/ml), 1ml IPTG(24mg /ml) and 2ml x-gal (20mg/ml) and mix uniformly.
      7. Paving plate (30-50ml/90mm petri dish).
      8.4 ℃ avoid light preservation.
       
      TB/Amp/ x-gal /IPTG flat medium
      Tryptone component concentration of 1.2% (W/V), 2.4% (W/V) Yeast extract, 0.4% (W/V) Glycerol, 17 M M KH2PO4, K2HPO4 72 mm, 0.1 mg/ml Ampicillin, 0.024 mg/ml IPTG, 0.04 mg/ml X - Gal, 1.5% (W/V) Agar.
      Dispensing quantity 1 l
      Preparation methods
      1. Prepared phosphate buffer (0.17 M KH2PO4, 0.72M K2HPO4) 100ml.
      Dissolve 2.31g KH2PO4 and 12.54g K2HPO4 in 90ml deionized water, stir and dissolve, add deionized water and set the capacity to 100ml, high temperature and high pressure sterilization.
      2. Weigh 12 g Tryptone, 24 g Yeast extract, 4 ml Glycerol into 1 l in the beaker.
      3. Add about 800ml of deionized water, stir well, dissolve and set the capacity to 1L, add 15g Agar, high pressure and high temperature sterilization.
      4. Stay cool solution to 60 ℃ below, add 100 ml of the sterilization phosphate buffer and 1 ml of Ampicillin (100 mg/ml), 1 ml IPTG (24 mg/ml) and 2 ml X - Gal (20 mg/ml).
      5. Planking (30-35ml medium /90mm dish).
      6. 4 ℃ after mixing avoid light preservation.
       
      Ampicillin (100 mg/ml)
      The component concentration is 100mg/ml Ampicillin
      Dispensing quantity 50 ml
      Preparation methods
      1. Weigh 5g Ampicillin and place it in a 50ml centrifuge tube.
      2. Add 40ml sterilized water, mix and dissolve thoroughly, and set the capacity to 50ml.
      3. Sterilize with 0.22um filter.
      4. Small packing (1 ml/copy), and 20 ℃.
       
      IPTG (24 mg/ml)
      The component concentration was 24mg/ml IPTG
      Dispensing quantity 50 ml
      Preparation methods
      1. Weigh 1g x-gal and place it in 50ml centrifugal tube.
      2. Add 4
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