- 定做培養(yǎng)基/定制培養(yǎng)基
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ATCC培養(yǎng)基2154,阿米巴蟲(chóng)LYI培養(yǎng)基
LYI Entamoeba Medium,ATCC medium 2154/ATCC medium:2154
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配套試劑
產(chǎn)品詳情
培養(yǎng)基配方（每升）
Sodium Chloride（氯化鈉） 1.00g
Yeast extract（酵母浸粉） 25.00g
Neutralized Liver Digest
（肝粉） 5.00g
L-Cysteine x HCl
（L-半胱氨酸鹽酸鹽） 1.00g
Ascorbic Acid（抗壞血酸） 0.20g
Ferric Ammonium Citrate
（檸檬酸鐵銨） 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-半胱氨酸鹽酸鹽） 1.00g
Ascorbic Acid（抗壞血酸） 0.20g
Ferric Ammonium Citrate
（檸檬酸鐵銨） 22.8mg
pH6.8±0.2（25℃）
使用說(shuō)明
稱(chēng)取本品32.23g于780ml去離子水中，加熱溶解，78ml/瓶分裝，121℃高壓滅菌15分鐘,冷卻，備用。
使用前，無(wú)菌操作每78ml培養(yǎng)基加入10ml葡萄糖緩沖液（10×）、10ml熱激活的成年牛血清和2ml Diamond’s Vitamin Solution，搖勻，備用。
使用前，無(wú)菌操作每78ml培養(yǎng)基加入10ml葡萄糖緩沖液（10×）、10ml熱激活的成年牛血清和2ml Diamond’s Vitamin Solution，搖勻，備用。
使用說(shuō)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.
使用說(shuō)明
Diamond維生素混合物107:
混合以上溶液，過(guò)濾除菌。-20°C避光保存。如果溶液呈現(xiàn)渾濁，應(yīng)該丟棄。渾濁出現(xiàn)說(shuō)明在配制過(guò)程中過(guò)量使用了氫氧化鈉。
水溶性維生素(Water-soluble Vitamins):
用75ml蒸餾水重懸核黃素，滴加 0.1N NaOH 溶液直至核黃素完全溶解。定容至100ml。用150ml沸騰的蒸餾水溶解煙酰胺和對(duì)氨基苯甲酸。用250ml溶解其余維生素。混合以上3種溶液。
生物素溶液(Biotin Solution):
用150ml蒸餾水重懸25mg生物素，滴加0.1 N NaOH溶液，直至生物素完全溶解。用蒸餾水定容至250ml。
葉酸溶液(Folic Acid Solution):
用150ml蒸餾水重懸25mg葉酸，滴加0.1 N NaOH溶液，直至葉酸完全溶解。用蒸餾水定容至250ml。
脂溶性維生素A、D、K
用63 ml 9.5%乙醇溶解維生素D2。然后加入維生素A。用300ml 5%吐溫80水溶液溶解維生素K�；旌蟽煞N溶液，用蒸餾水定容至3000ml。
維生素E溶液(Vitamin E Solution):
| 水溶性維生素溶液 | 500.0 ml |
| 生物素溶液 | 250.0 ml |
| 葉酸溶液 | 250.0 ml |
| 脂溶性維生素A、D、K | 2500.0 ml |
| 維生素E溶液 | 250.0 ml |
混合以上溶液，過(guò)濾除菌。-20°C避光保存。如果溶液呈現(xiàn)渾濁，應(yīng)該丟棄。渾濁出現(xiàn)說(shuō)明在配制過(guò)程中過(guò)量使用了氫氧化鈉。
水溶性維生素(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 |
| 蒸餾水 | 500 ml |
用75ml蒸餾水重懸核黃素，滴加 0.1N NaOH 溶液直至核黃素完全溶解。定容至100ml。用150ml沸騰的蒸餾水溶解煙酰胺和對(duì)氨基苯甲酸。用250ml溶解其余維生素。混合以上3種溶液。
生物素溶液(Biotin Solution):
| 生物素 | 25.0 mg |
| 蒸餾水 | 250 ml |
用150ml蒸餾水重懸25mg生物素，滴加0.1 N NaOH溶液，直至生物素完全溶解。用蒸餾水定容至250ml。
葉酸溶液(Folic Acid Solution):
| 葉酸 | 25.0 mg |
| 蒸餾水 | 250 ml |
用150ml蒸餾水重懸25mg葉酸，滴加0.1 N NaOH溶液，直至葉酸完全溶解。用蒸餾水定容至250ml。
脂溶性維生素A、D、K
| 維生素D2 | 300.0 mg |
| 維生素A | 300.0 mg |
| 維生素K | 60.0 mg |
| 9.5%乙醇 | 63 ml |
| 5%吐溫80水溶液 | 300 ml |
| 蒸餾水定容至 | 3000 ml |
用63 ml 9.5%乙醇溶解維生素D2。然后加入維生素A。用300ml 5%吐溫80水溶液溶解維生素K�；旌蟽煞N溶液，用蒸餾水定容至3000ml。
維生素E溶液(Vitamin E Solution):
| 維生素E | 25.0 mg |
| 蒸餾水 |
250.0 ml |
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