CN1255424C - 注射用人胎盘核糖核酸的制备方法 - Google Patents
注射用人胎盘核糖核酸的制备方法 Download PDFInfo
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- CN1255424C CN1255424C CNB200310109446XA CN200310109446A CN1255424C CN 1255424 C CN1255424 C CN 1255424C CN B200310109446X A CNB200310109446X A CN B200310109446XA CN 200310109446 A CN200310109446 A CN 200310109446A CN 1255424 C CN1255424 C CN 1255424C
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Abstract
Description
名称 | 参数 | ||||
配置试剂步骤 | |||||
a操作中 | 十二烷基硫酸钠(重量比) | 0.4% | 0.45% | 0.55% | 0.6% |
聚乙烯硫酸酯(重量比) | 0.1% | 0.15% | 0.25% | 0.3% | |
乙二胺四乙酸二钠(重量比) | 0.1% | 0.15% | 0.25% | 0.3% | |
b操作中 | 振摇(次) | 200 | 250 | 350 | 400 |
避光静置(小时) | 8 | 10 | 13 | 14 | |
d操作中 | 醋酸钾(重量比) | 6% | 6.5% | 7.5% | 8% |
e操作中 | 醋酸钾(重量比) | 2% | 2.5% | 3.5% | 4% |
聚乙烯硫酸酯(重量比) | 0.1% | 0.15% | 0.25% | 0.3% | |
破碎粗提步骤 | |||||
b操作中 | 酚饱和缓冲液(重量倍) | 4 | 4.5 | 5.5 | 6 |
缓冲液饱和酚(重量倍) | 4 | 4.5 | 5.5 | 6 | |
转速(转/分钟) | 10000 | 10500 | 11500 | 12000 | |
捣碎(分钟) | 1 | 1.25 | 1.75 | 2 | |
停(分钟) | 2 | 3 | 4 | 5 | |
捣碎、停(次) | 3 | 3 | 2 | 2 | |
c操作中 | 水浴温度(℃) | 50 | 60 | 70 | 80 |
搅拌(分钟) | 10 | 12 | 18 | 20 | |
盐水浴温度(℃) | -20 | -18 | -12 | -10 | |
速冷至温度(℃) | 0 | 3 | 8 | 10 | |
精提分离步骤 | |||||
a操作中 | 冰箱温度(℃) | 0 | 3 | 8 | 10 |
b操作中 | 酚饱和缓冲液(重量倍) | 1/2 | 4/7 | 5/8 | 2/3 |
搅拌(分钟) | 10 | 12 | 14 | 15 | |
c操作中 | 缓冲液饱和酚(重量倍) | 1/2 | 4/7 | 5/8 | 2/3 |
搅拌(分钟) | 10 | 12 | 14 | 15 | |
d操作中 | 醋酸钾(重量比) | 5% | 6% | 7% | 8% |
无水乙醇(重量倍) | 2 | 2.25 | 2.75 | 3 | |
无水乙醇温度(℃) | -10 | -12 | -18 | -20 | |
静置(小时) | 8 | 10 | 12 | 14 | |
纯化步骤 | |||||
a操作中 | 0.3mol/l醋酸盐缓冲液(ml) | 50 | 60 | 70 | 80 |
无水乙醇(重量倍) | 2 | 2.25 | 2.75 | 3 | |
无水乙醇温度(℃) | -10 | -12 | -18 | -20 |
静置(小时) | 1 | 1.25 | 1.75 | 2 |
b操作中0.1mol/l醋酸盐洗液(重量倍) | 6 | 6.5 | 7.5 | 8 |
c操作中0.1mol/l醋酸盐洗液(重量倍) | 6 | 6.5 | 7.5 | 8 |
配制注射用人胎盘核糖核酸的步骤 | ||||
核糖核酸(mg) | 10 | 20 | 40 | 50 |
甘露醇(mg) | 100 | 200 | 300 | 400 |
右旋糖苷(mg) | 0 | 15 | 35 | 50 |
序号 | 指标名称 | 热酚法提取的RNA | 冷酚法提取的RNA |
1 | 性状 | 无色透明液体 | 无色透明液体 |
2 | PH值 | 6.2 | 6.4 |
3 | 地衣酚试验 | 兰绿色 | 兰绿色 |
4 | 紫外分光光度计测定(样品100倍稀释) | ||
OD260/OD230 | 1.690 | 1.625 | |
OD260/OD280 | 2.51 | 2.24 | |
5 | RNA含量(mg/ml) | 10.0 | 7.5 |
6 | 蛋白含量(mg/ml) | 0.02375 | 0.01650 |
7 | 生物活性测定(E.RFC试验) | 40.0% | 38.0% |
Claims (6)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
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CNB200310109446XA CN1255424C (zh) | 2003-12-16 | 2003-12-16 | 注射用人胎盘核糖核酸的制备方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
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CNB200310109446XA CN1255424C (zh) | 2003-12-16 | 2003-12-16 | 注射用人胎盘核糖核酸的制备方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN1580068A CN1580068A (zh) | 2005-02-16 |
CN1255424C true CN1255424C (zh) | 2006-05-10 |
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Application Number | Title | Priority Date | Filing Date |
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CNB200310109446XA Expired - Lifetime CN1255424C (zh) | 2003-12-16 | 2003-12-16 | 注射用人胎盘核糖核酸的制备方法 |
Country Status (1)
Country | Link |
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CN (1) | CN1255424C (zh) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102321614A (zh) * | 2011-09-09 | 2012-01-18 | 杭州华津药业股份有限公司 | 从动物加工副产物中提取rna活性物的方法 |
CN106065400B (zh) * | 2015-04-24 | 2019-12-06 | 上海晟燃生物科技有限公司 | 一种核糖核酸保护剂、试剂盒、应用及保存方法 |
-
2003
- 2003-12-16 CN CNB200310109446XA patent/CN1255424C/zh not_active Expired - Lifetime
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CN1580068A (zh) | 2005-02-16 |
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