计算溶液所需的质量、体积或浓度。
为什么选择此级别
，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
2-8°C储存,室温。低温运输 。请查阅批次 COA 获取详细规格。
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文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
产品内容
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产品简介
本试剂盒适用于从新鲜或冷冻的血清、血浆、淋巴液等无细胞体液中提取游离DNA。本试剂盒采用可以特异性结合核酸的离心吸附柱和独特的缓冲液系统，样品裂解后，游离DNA在高盐条件下与硅胶膜结合，在低盐、高pH值时游离DNA从硅胶膜上洗脱下来。本品可以处理0.1-1 ml的液体样本，配置的高效微量吸附柱洗脱体积可低至20 μl。纯化的DNA产量高、质量好，最大限度去除蛋白、色素、脂类及其他抑制物， 游离DNA得率与样品的类型，储存条件、时间以及个体间差异有很大关系。纯化得到的游离DNA质量稳定、可靠，可直接用于PCR、荧光定量PCR和二代测序等分子生物学实验。
自备试剂：无水乙醇、异丙醇。
实验前准备及重要注意事项
1. 向Proteinase K中加入5 ml的Proteinase K Storage Buffer使其溶解，-20℃保存。配制好的Proteinase K勿长时间室温放置。
2. 样品应避免反复冻融，否则会导致提取量下降。
3. 本试剂盒可以提取0.1-1 ml液体样品。
4. 使用前请检查Buffer CL、Buffer CB是否出现结晶或者沉淀，如有结晶或者沉淀，请将Buffer CL、Buffer CB于56℃水浴孵育重新溶解。
5. 第一次使用前应按照试剂瓶标签的说明先在Buffer CB中加入异丙醇，混合均匀， 并在试剂瓶标签上做好标记。
6. 第一次使用前应按照试剂瓶标签的说明先在Buffer GW1和Buffer GW2中加入无水乙醇，混合均匀，并在试剂瓶标签上做好标记。
7. 实验开始前请将水浴锅预热至60℃。
8. 可将洗脱缓冲液Buffer EBL预热至60℃后使用。
操作步骤
1. 向离心管（自备）中加入20 μl Proteinase K。
2. 加入200 μl血清/血浆样本。
注意：当样品量超过200 μl时，请等比例增加Proteinase K、Buffer CL和Buffer CB试剂用量，具体试剂加入量可参考附表。
3. 加入160 μl Buffer CL，颠倒混匀，剧烈震荡至少30秒。
4．60℃孵育30分钟，其间颠倒混匀数次。
注意：200 μl 血清/血浆样本60℃孵育10-15分钟即可。
5. 加入360 μl Buffer CB（使用前检查是否加入异丙醇），震荡至彻底混匀。
6. 冰浴5分钟，短暂离心，使管壁和壁盖上的液体集中到管底。
7. 将步骤6所得溶液全部加入到已装入收集管的吸附柱（Spin Columns DF）中，若一次不能加完溶液，可分多次转入。12,000 rpm离心1分钟，倒掉收集管中的废液，将吸附柱重新放回收集管中。
8. 向吸附柱中加入500 μl Buffer GW1（使用前检查是否加入无水乙醇）, 12,000 rpm 离心30秒，倒掉收集管中的废液，将吸附柱重新放回收集管中。
9. 向吸附柱中加入750 μl Buffer GW2（使用前检查是否加入无水乙醇），12,000 rpm 离心30秒，倒掉收集管中的废液，将吸附柱重新放回收集管中。
10. 向吸附柱中加入750 μl 无水乙醇，12,000 rpm离心30秒，倒掉收集管中的废液，将吸附柱重新放回收集管中。
11.12,000 rpm离心2分钟，倒掉收集管中的废液。将吸附柱置于室温数分钟，以彻底晾干。
注意：这一步的目的是将吸附柱中残余的乙醇去除，乙醇的残留会影响后续的酶促反应。
12. 将吸附柱置于新的离心管中，向吸附柱的中间部位悬空加入20-100 μl Buffer EBL或灭菌水，室温放置2-5分钟，12,000 rpm离心1分钟，收集DNA溶液，-20℃保存DNA。
注意：1）如果下游实验对pH值敏感，可以用灭菌水洗脱。洗脱液的pH值对洗脱效率有很大影响， 若用水做洗脱液应保证其pH值在7.0-8.5（可以用NaOH将水的pH值调到此范围），pH值低于7.0 时洗脱效率不高。
2） 将洗脱缓冲液Buffer EBL预热至60℃后使用,且离心之前室温孵育5分钟，可以增加产量。
3） 如果要提高DNA的终浓度，可以将得到的溶液重新加入到吸附柱中，室温放置2-5分钟，12,000 rpm 离 心 1 分 钟 。
4）因为保存在水中的DNA会受到酸性水解作用的影响，如需长期保存，推荐用Buffer EBL洗脱并于-20℃保存。
附表：不同样本量推荐试剂加入量
Product content
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Product Introduction
This kit is suitable for the extraction of free DNA from fresh or frozen serum, plasma, lymph fluid and other cell-free body fluids.This kit adopts centrifugal adsorption columns that can specifically bind nucleic acids and a unique buffer system.After the sample is lysed, the free DNA binds to the silica gel membrane under high salt conditions, and the free DNA elutes from the silica gel membrane at low salt and high pH. The product can handle liquid samples of 0.1-1 ml, and the elution volume of the configured high-efficiency micro adsorption column can be as low as 20 μl. The purified DNA is of high yield and quality, with maximum removal of proteins, pigments, lipids, and other inhibitors, and the rate of free DNA yield is highly dependent on the type of samples, storage conditions, time, and inter-individual variations. The quality of free DNA obtained from purification is stable and reliable, and can be directly used in molecular biology experiments such as PCR, fluorescence quantitative PCR and second generation sequencing.
Self-contained reagents: anhydrous ethanol, isopropanol.
Pre-experiment Preparation and Important Notes
Add 5 ml of Proteinase K Storage Buffer to Proteinase K to dissolve it and store it at -20℃. Do not leave the prepared Proteinase K at room temperature for a long time.
Repeated freezing and thawing of the sample should be avoided, as this can lead to a decrease in extraction.
This kit can extract 0.1-1 ml of liquid samples.
Before use, please check Buffer CL, Buffer CB for crystallization or precipitation, if there is any crystallization or precipitation, please re-dissolve Buffer CL, Buffer CB by incubation at 56℃ in a water bath.
Before first use isopropyl alcohol should be added to Buffer CB according to the instructions on the reagent bottle label, mixed well, and labeled on the reagent bottle label.
Before the first use, anhydrous ethanol should be added to Buffer GW1 and Buffer GW2 according to the instructions on the label of the reagent bottle, mixed well, and labeled on the label of the reagent bottle.
Preheat the water bath to 60°C before starting the experiment.
The elution buffer Buffer EBL can be preheated to 60°C and used.
Operation steps
Add 20 μl of Proteinase K to the centrifuge tube (supplied).
Add 200 μl of serum/plasma sample.
Note: When the sample volume exceeds 200 μl, please increase the amount of Proteinase K, Buffer CL and Buffer CB reagents in equal proportions, and the specific amount of reagents added can be referred to the attached table.
3. Add 160 μl Buffer CL, mix upside down and shake vigorously for at least 30 seconds.
4. Incubate at 60°C for 30 minutes, during which time mixing was inverted several times.
Note: Incubation of 200μl serum/plasma samples at 60°C for 10-15 minutes is sufficient.
Add 360 μl of Buffer CB (check for addition of isopropanol before use) and shake until thoroughly mixed.
Ice bath for 5 minutes and centrifuge briefly to concentrate the liquid on the walls and wall caps to the bottom of the tube.
Add all of the solution obtained in step 6 to the adsorption columns (Spin Columns DF) that have been loaded into the collection tubes, and if the solution cannot be added all at once, it can be transferred in several times. centrifuge the columns at 12,000 rpm for 1 minute, pour off the waste solution from the collection tubes, and put the columns back into the collection tubes.
Add 500μl of Buffer GW1 to the adsorbent column (check that anhydrous ethanol is added before use),centrifuge the column at 12,000rpm for 30 seconds, pour off the waste liquid in the collection tube, and put the adsorbent column back into the collection tube.
Add 750 μl Buffer GW2 to the adsorbent column (check that anhydrous ethanol is added before use), centrifuge at 12,000 rpm for 30 seconds, pour off the waste liquid in the collection tube, and put the adsorbent column back into the collection tube.
10. Add 750 μl of anhydrous ethanol to the adsorbent column and centrifuge at 12,000 rpm for 30 s. Pour off the waste liquid in the collection tube and put the adsorbent column back into the collection tube.
11. Centrifuge at 12,000 rpm for 2 minutes and pour off the waste liquid in the collection tube. Leave the adsorption column at room temperature for several minutes to dry thoroughly.
Note: The purpose of this step is to remove residual ethanol from the adsorption column, which can interfere with the subsequent enzymatic reaction.
12. Place the adsorption column in a new centrifuge tube, add 20-100 μl Buffer EBL or sterilized water to the middle part of the adsorption column overhanging the column, leave it at room temperature for 2-5 minutes, centrifuge at 12,000 rpm for 1 minute, collect the DNA solution, and store the DNA at -20℃.
Note: 1) If the downstream experiment is sensitive to pH, you can use sterilized water for elution. The pH value of the eluent has a great influence on the elution efficiency, if water is used as the eluent should ensure that its pH value is 7.0-8.5 (you can use NaOH to adjust the pH value of water to this range), and the elution efficiency is not high when the pH value is lower than 7.0.
2) Preheat the elution buffer BufferEBL to 60℃ and use it, and incubate it at room temperature for 5 minutes before centrifugation to increase the yield.
3) If the final concentration of DNA is to be increased, the resulting solution can be reintroduced into the adsorption column and left at room temperature for 2-5 minutes and centrifuged at 12,000 rpm for 1 minute.
4) Because DNA preserved in water will be affected by acidic hydrolysis, for long-term storage, it is recommended to elute it with Buffer EBL and store it at -20℃.
Table: Recommended reagent additions for different sample sizes
规格
英文名称
CWhipro Circulating Nucleic Acid Kit
储存与运输
储存条件
2-8°C储存,室温
运输条件
低温运输
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
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技术文档和文章
溶液计算器
常见问题
本产品应如何储存？
请于2–8 °C条件下保存。需冷藏保存以维持规定的有效期。
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