- 定做培养基/定制培养基
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ATCC培养基2154,阿米巴虫LYI培养基
LYI Entamoeba Medium,ATCC medium 2154/ATCC medium:2154
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配套试剂
产品详情
培养基配方（每升）
Sodium Chloride（氯化钠） 1.00g
Yeast extract（酵母浸粉） 25.00g
Neutralized Liver Digest
（肝粉） 5.00g
L-Cysteine x HCl
（L-半胱氨酸盐酸盐） 1.00g
Ascorbic Acid（抗坏血酸） 0.20g
Ferric Ammonium Citrate
（柠檬酸铁铵） 22.8mg
pH6.8±0.2（25℃）
Yeast extract（酵母浸粉） 25.00g
Neutralized Liver Digest
（肝粉） 5.00g
L-Cysteine x HCl
（L-半胱氨酸盐酸盐） 1.00g
Ascorbic Acid（抗坏血酸） 0.20g
Ferric Ammonium Citrate
（柠檬酸铁铵） 22.8mg
pH6.8±0.2（25℃）
使用说明
称取本品32.23g于780ml去离子水中，加热溶解，78ml/瓶分装，121℃高压灭菌15分钟,冷却，备用。
使用前，无菌操作每78ml培养基加入10ml葡萄糖缓冲液（10×）、10ml热激活的成年牛血清和2ml Diamond’s Vitamin Solution，摇匀，备用。
使用前，无菌操作每78ml培养基加入10ml葡萄糖缓冲液（10×）、10ml热激活的成年牛血清和2ml Diamond’s Vitamin Solution，摇匀，备用。
使用说明
Diamond's Vitamin Mixture (107)
Water-soluble B Vitamins (see below)........................500.0 ml
Biotin Solution (see below).........................................250.0 ml
Folic Acid Solution (see below)..................................250.0 ml
Lipid-soluble Vitamins A, D, and K (see below)…….2,500.0 ml
Vitamin E Solution (see below)..................................250.0 ml
Sterilize final solution by passage through a 0.22 micron filter. Store the
completed, clear mixture at -22C. Thaw and warm to room temperature before
use.
If the mixture appears turbid, it should be discarded. Development of turbidity is
an indication that an excess of NaOH has been used in the preparation of one of
the stock solutions.J. Parasitol. 54: 1047-1056.
Water-soluble Vitamins:
Water Solution A (see below)...........150.0 ml
Water Solution B (see below)...........150.0 ml
Water Solution C (see below)...........100.0 ml
Distilled water to................................100.0 ml
Combine solutions A, B, and C and bring total volume to 500 ml with distilled
water.
Water Solution A:
Niacin......................................................62.5 mg
p-Aminobenzoic acid...............................12.5 mg
Distilled water..........................................150.0 ml
Dissolve solid ingredients in boiling distilled water; restore final volume to 150 ml.
Water Solution B:
Niacinamide............................................62.5 mg
Pyridoxine hydrochloride........................62.5 mg
Pyridoxal hydrochloride..........................62.5 mg
Thiamine hydrochloride..........................25.0 mg
Calcium pantothenate............................25.0 mg
i-Inositol.................................................125.0 mg
Choline chloride.....................................1250.0 mg
Distilled water to....................................150.0 ml
Dissolve solid ingredients in 125 ml of the water. Bring final volume to 150 ml.
Water Solution C:
Riboflavin...............................................25.0 mg
2154
0.1 N NaOH (as needed)
Distilled water to..................................100.0 ml
Add riboflavin to 75 ml of the distilled water; add NaOH drop by drop until the
riboflavin dissolves. Bring total volume to 100 ml with distilled water.
Biotin Solution:
Biotin...................................................30.0 mg
0.1 N NaOH (as needed)
Distilled water to..................................300.0 ml
Add biotin to 200 ml of distilled water; add NaOH drop by drop until the
biotin dissolves. Bring total volume to 300 ml with distilled water.
Folic Acid Solution:
Folic acid..............................................30.0 mg
0.1 N NaOH (as needed)
Distilled water to..................................300.0 ml
Add folic acid to 200 ml of distilled water; add NaOH drop by drop until
the folic acid dissolves. Bring total volume to 300 ml with distilled
water.
Lipid-soluble Vitamins A, D, and K:
Lipid Solution A (see below)
Lipid Solution B (see below)
Distilled water to..................................3000.0 L
Combine Lipid Solution B with Lipid Solution A. Add distilled water to
bring volume to 3000 ml.
Lipid Solution A:
Vitamin D2 (caciferol).........................300.0 mg
Ethyl alcohol 9.5% (v/v)…………........63.0 ml
Vitamin A (crystalline alcohol)……….300.0 mg
Dissolve vitamin D2 in ethyl alcohol; add vitamin A.
Lipid Solution B:
Vitamin K (menadione sodium bisulfite)........60.0 mg
Tween 80 aqueous solution 5% (v/v).............300.0 ml
Dissolve vitamin K in the Tween 80 solution.
Lipid Solution C:
Vitamin E (alpha tocopherol acetate)..........25.0 mg
2154
Distilled water...........................................250.0 ml
Dissolve vitamin E in distilled water.
Water-soluble B Vitamins (see below)........................500.0 ml
Biotin Solution (see below).........................................250.0 ml
Folic Acid Solution (see below)..................................250.0 ml
Lipid-soluble Vitamins A, D, and K (see below)…….2,500.0 ml
Vitamin E Solution (see below)..................................250.0 ml
Sterilize final solution by passage through a 0.22 micron filter. Store the
completed, clear mixture at -22C. Thaw and warm to room temperature before
use.
If the mixture appears turbid, it should be discarded. Development of turbidity is
an indication that an excess of NaOH has been used in the preparation of one of
the stock solutions.J. Parasitol. 54: 1047-1056.
Water-soluble Vitamins:
Water Solution A (see below)...........150.0 ml
Water Solution B (see below)...........150.0 ml
Water Solution C (see below)...........100.0 ml
Distilled water to................................100.0 ml
Combine solutions A, B, and C and bring total volume to 500 ml with distilled
water.
Water Solution A:
Niacin......................................................62.5 mg
p-Aminobenzoic acid...............................12.5 mg
Distilled water..........................................150.0 ml
Dissolve solid ingredients in boiling distilled water; restore final volume to 150 ml.
Water Solution B:
Niacinamide............................................62.5 mg
Pyridoxine hydrochloride........................62.5 mg
Pyridoxal hydrochloride..........................62.5 mg
Thiamine hydrochloride..........................25.0 mg
Calcium pantothenate............................25.0 mg
i-Inositol.................................................125.0 mg
Choline chloride.....................................1250.0 mg
Distilled water to....................................150.0 ml
Dissolve solid ingredients in 125 ml of the water. Bring final volume to 150 ml.
Water Solution C:
Riboflavin...............................................25.0 mg
2154
0.1 N NaOH (as needed)
Distilled water to..................................100.0 ml
Add riboflavin to 75 ml of the distilled water; add NaOH drop by drop until the
riboflavin dissolves. Bring total volume to 100 ml with distilled water.
Biotin Solution:
Biotin...................................................30.0 mg
0.1 N NaOH (as needed)
Distilled water to..................................300.0 ml
Add biotin to 200 ml of distilled water; add NaOH drop by drop until the
biotin dissolves. Bring total volume to 300 ml with distilled water.
Folic Acid Solution:
Folic acid..............................................30.0 mg
0.1 N NaOH (as needed)
Distilled water to..................................300.0 ml
Add folic acid to 200 ml of distilled water; add NaOH drop by drop until
the folic acid dissolves. Bring total volume to 300 ml with distilled
water.
Lipid-soluble Vitamins A, D, and K:
Lipid Solution A (see below)
Lipid Solution B (see below)
Distilled water to..................................3000.0 L
Combine Lipid Solution B with Lipid Solution A. Add distilled water to
bring volume to 3000 ml.
Lipid Solution A:
Vitamin D2 (caciferol).........................300.0 mg
Ethyl alcohol 9.5% (v/v)…………........63.0 ml
Vitamin A (crystalline alcohol)……….300.0 mg
Dissolve vitamin D2 in ethyl alcohol; add vitamin A.
Lipid Solution B:
Vitamin K (menadione sodium bisulfite)........60.0 mg
Tween 80 aqueous solution 5% (v/v).............300.0 ml
Dissolve vitamin K in the Tween 80 solution.
Lipid Solution C:
Vitamin E (alpha tocopherol acetate)..........25.0 mg
2154
Distilled water...........................................250.0 ml
Dissolve vitamin E in distilled water.
使用说明
Diamond维生素混合物107:
混合以上溶液，过滤除菌。-20°C避光保存。如果溶液呈现浑浊，应该丢弃。浑浊出现说明在配制过程中过量使用了氢氧化钠。
水溶性维生素(Water-soluble Vitamins):
用75ml蒸馏水重悬核黄素，滴加 0.1N NaOH 溶液直至核黄素完全溶解。定容至100ml。用150ml沸腾的蒸馏水溶解烟酰胺和对氨基苯甲酸。用250ml溶解其余维生素。混合以上3种溶液。
生物素溶液(Biotin Solution):
用150ml蒸馏水重悬25mg生物素，滴加0.1 N NaOH溶液，直至生物素完全溶解。用蒸馏水定容至250ml。
叶酸溶液(Folic Acid Solution):
用150ml蒸馏水重悬25mg叶酸，滴加0.1 N NaOH溶液，直至叶酸完全溶解。用蒸馏水定容至250ml。
脂溶性维生素A、D、K
用63 ml 9.5%乙醇溶解维生素D2。然后加入维生素A。用300ml 5%吐温80水溶液溶解维生素K。混合两种溶液，用蒸馏水定容至3000ml。
维生素E溶液(Vitamin E Solution):
| 水溶性维生素溶液 | 500.0 ml |
| 生物素溶液 | 250.0 ml |
| 叶酸溶液 | 250.0 ml |
| 脂溶性维生素A、D、K | 2500.0 ml |
| 维生素E溶液 | 250.0 ml |
混合以上溶液，过滤除菌。-20°C避光保存。如果溶液呈现浑浊，应该丢弃。浑浊出现说明在配制过程中过量使用了氢氧化钠。
水溶性维生素(Water-soluble Vitamins):
| Niacin | 62.5 mg |
| p-Aminobenzoic acid | 12.5 mg |
| Niacinamide | 62.5 mg |
| Pyridoxine hydrochloride | 62.5 mg |
| Pyridoxal hydrochloride | 62.5 mg |
| Thiamine hydrochloride | 25.0 mg |
| Calcium pantothenate | 25.0 mg |
| i-Inositol | 125.0 mg |
| Choline chloride | 1250.0 mg |
| Riboflavin | 25.0 mg |
| 蒸馏水 | 500 ml |
用75ml蒸馏水重悬核黄素，滴加 0.1N NaOH 溶液直至核黄素完全溶解。定容至100ml。用150ml沸腾的蒸馏水溶解烟酰胺和对氨基苯甲酸。用250ml溶解其余维生素。混合以上3种溶液。
生物素溶液(Biotin Solution):
| 生物素 | 25.0 mg |
| 蒸馏水 | 250 ml |
用150ml蒸馏水重悬25mg生物素，滴加0.1 N NaOH溶液，直至生物素完全溶解。用蒸馏水定容至250ml。
叶酸溶液(Folic Acid Solution):
| 叶酸 | 25.0 mg |
| 蒸馏水 | 250 ml |
用150ml蒸馏水重悬25mg叶酸，滴加0.1 N NaOH溶液，直至叶酸完全溶解。用蒸馏水定容至250ml。
脂溶性维生素A、D、K
| 维生素D2 | 300.0 mg |
| 维生素A | 300.0 mg |
| 维生素K | 60.0 mg |
| 9.5%乙醇 | 63 ml |
| 5%吐温80水溶液 | 300 ml |
| 蒸馏水定容至 | 3000 ml |
用63 ml 9.5%乙醇溶解维生素D2。然后加入维生素A。用300ml 5%吐温80水溶液溶解维生素K。混合两种溶液，用蒸馏水定容至3000ml。
维生素E溶液(Vitamin E Solution):
| 维生素E | 25.0 mg |
| 蒸馏水 |
250.0 ml |
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