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
室温。常规运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
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：
本试剂盒采用新型硅基质膜技术和试剂配方，通过快速简单的结合-洗涤-洗脱三 步即可从PCR产物或酶反应液（酶切，连接，探针标记等）中纯化回收100 bp-10 kb的 DNA片段，每个吸附柱最高可吸附10 μg的DNA，同时最大限度的去除引物、寡核苷酸、 酶等杂质。纯化回收的DNA纯度及浓度高，完整性好，回收率高，可直接用于测序、 连接和转化、标记、体外转录等分子生物学实验。
自备试剂：无水乙醇。
实验前准备及重要注意事项：
1 ．所有组分可在干燥、室温（15-30℃)环境稳定保存1年，更长时间保存可置于2- 8℃。 当溶液低温保存时,使用前需在室温中放置一段时间，恢复至室温后使用。
2 ．本试剂盒可无选择性回收溶液中所有DNA片段，如需选择性回收特定片段，同时去除其他不同大小片段，请选择我公司的胶回收试剂盒。
3 ．第一次使用前应按照试剂瓶标签的说明在Buffer PW中加入无水乙醇。
4 ．使用前请检查Buffer PB是否出现结晶或者沉淀，如有结晶或者沉淀现象，可在37℃ 水浴几分钟，即可恢复澄清。
5 ．回收效率与初始DNA量和洗脱体积有关，初始量越少，洗脱体积越少，回收率越低。
6 ．所有离心步骤均可室温下进行。
操作步骤
1.估计DNA反应液的体积，加入5倍体积的 Buffer PB，充分混匀（无需去除石蜡油或 矿物油） 。
注意：1) 如DNA反应体系为50 µl(不包括石蜡油体积)，则加入250 µl Buffer PB。
2)在加入Buffer PB后检测溶液的pH值，若pH值＞7.5 ，可向其中加入10-30 μl的3 M醋酸钠（pH5.0 ）， 从而将pH值调到5-7。
2.柱平衡：向已装入收集管中的吸附柱（Spin Columns DM）中加入200 μl Buffer PS， 13,000 rpm（~16,200×g）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。
3.将步骤1中所得溶液加入到已装入收集管的吸附柱中，室温放置1分钟， 13,000 rpm 离心30-60 s，倒掉收集管中的废液，将吸附柱放回收集管中。
注意：吸附柱容积为750 µl，若样品体积大于750 µl可分批加入 。
4.向吸附柱中加入500 µl Buffer PW（使用前请先检查是否已加入无水乙醇）13,000 rpm 离心30-60 s，倒掉收集管中的废液，将吸附柱放回收集管中。
注意：如果纯化的DNA用于盐敏感实验（例如平末端连接实验或直接测序），建议加入Buffer PW后 静置2-5分钟再离心。
5.13,000 rpm离心1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（酶切、 PCR等）。为确保下游实验不受残留乙醇的影响，建议将吸附柱开盖，置于室温放置数分钟，以彻
底晾干吸附材料中残余的乙醇。
6.将吸附柱放到一个新离心管（自备）中，向吸附膜中间位置悬空滴加30-50 µl Buffer EB，室温放置1分钟。 13,000 rpm离心1分钟，收集DNA溶液。 -20℃保存DNA。
注意：
1 ）洗脱液的pH值对于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之间 （可以用 NaOH将水的pH值调到此范围）。
2 ）为了提高DNA的回收量，可将离心得到的溶液重新加到吸附柱中，室温放置2分钟，13,000 rpm 离心1分钟。
3 ）洗脱体积不应小于30 μl ，体积过少会影响回收效率。
4 ）回收＞ 10 kb的DNA片段时， Buffer EB应在50 ℃水浴中预热，适当延长吸附和洗脱时间，可增加 回收效率。
Product content:
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Product Introduction:
This reagent kit adopts a new silicon-based membrane technology and reagent formula. Through a rapid and simple three-step process of binding, washing, and elution, 100 bp-10 kb DNA fragments can be purified and recovered from PCR products or enzyme reaction solutions (enzyme cutting, linking, probe labeling, etc.). Each adsorption column can adsorb up to 10 kb of DNA fragments μ G DNA, while minimizing impurities such as primers, oligonucleotides, enzymes, etc. The purified and recovered DNA has high purity and concentration, good integrity, and high recovery rate, and can be directly used for molecular biology experiments such as sequencing, linking and transformation, labeling, and in vitro transcription.
Self prepared reagent: anhydrous ethanol.
Preparation and important precautions before the experiment:
1. All components can be stably stored in a dry, room temperature (15-30 ℃) environment for 1 year, and can be stored at 2-8 ℃ for longer periods of time. When the solution is stored at low temperature, it should be left at room temperature for a period of time before use, and then restored to room temperature before use.
2. This reagent kit can selectively recover all DNA fragments from the solution. If you need to selectively recover specific fragments while removing other fragments of different sizes, please choose our company's gel recovery reagent kit.
3.Before the first use, anhydrous ethanol should be added to the Buffer PW according to the instructions on the reagent bottle label.
4. Before use, please check if there is any crystallization or precipitation in the Buffer PB. If there is any crystallization or precipitation, you can take a water bath at 37 ℃ for a few minutes to restore clarity.
5. The recovery efficiency is related to the initial amount of DNA and the elution volume. The smaller the initial amount, the smaller the elution volume, and the lower the recovery rate.
6. All centrifugation steps can be performed at room temperature.
Operation steps:
1. Estimate the volume of DNA reaction solution, add 5 times the volume of Buffer PB, and mix thoroughly (without removing paraffin or mineral oil).
Note:
1) If the DNA reaction system is 50 µ l (excluding paraffin oil volume), add 250 µ l Buffer PB.
2) After adding Buffer PB, check the pH value of the solution. If the pH value is greater than 7.5, add 10-30 to it μ 3 M sodium acetate (pH 5.0) was used to adjust the pH value to 5-7.
2. Column balance: Add 200 to the spin columns DM that have been loaded into the collection tube μ Centrifuge at 13000 rpm (~16200 × g) for 1 minute, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.
3. Add the solution obtained in step 1 to the adsorption column that has been loaded into the collection tube, let it stand at room temperature for 1 minute, centrifuge at 13000 rpm for 30-60 seconds, discard the waste liquid in the collection tube, and place the adsorption column in the collection tube.
Attention: The volume of the adsorption column is 750 µ l. If the sample volume is greater than 750 µ l, it can be added in batches.
4. Add 500 µ l of Buffer PW to the adsorption column (please check if anhydrous ethanol has been added before use), centrifuge at 13000 rpm for 30-60 seconds, discard the waste liquid in the collection tube, and place the adsorption column in the recovery tube.
Note: If purified DNA is used for salt sensitivity experiments (such as flat end ligation experiments or direct sequencing), it is recommended to add Buffer PW and let it stand for 2-5 minutes before centrifugation.
5.13000 rpm for 1 minute and discard the waste liquid from the collection tube. Place the adsorption column at room temperature for a few minutes to thoroughly air dry.
Note: The purpose of this step is to remove residual ethanol from the adsorption column, which can affect subsequent enzymatic reactions (such as enzyme digestion, PCR, etc.). To ensure that downstream experiments are not affected by residual ethanol, it is recommended to open the cover of the adsorption column and place it at room temperature for a few minutes to thoroughly dry the residual ethanol in the adsorbent material at the bottom.
6. Place the adsorption column into a new centrifuge tube (provided by oneself), add 30-50 µ l Buffer EB to the middle position of the adsorption membrane by hanging droplets, and let it stand at room temperature for 1 minute. Centrifuge at 13000 rpm for 1 minute and collect DNA solution- Store DNA at 20 ℃.
Attention:
1) The pH value of the eluent has a significant impact on the elution efficiency. If water is used as the eluent, its pH value should be ensured to be between 7.0-8.5 (the pH value of water can be adjusted to this range using NaOH).
2) To improve the recovery of DNA, the solution obtained by centrifugation can be added back to the adsorption column, left at room temperature for 2 minutes, and centrifuged at 13000 rpm for 1 minute.
3) The elution volume should not be less than 30 μ l. A small volume will affect the recovery efficiency.
规格
英文名称
DNA Clean-up Kit
储存与运输
储存条件
室温
运输条件
常规运输
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
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溶液计算器
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