计算溶液所需的质量、体积或浓度。
BioReagent,分子生物学级 ,适用于对基线干扰要求严格的色谱和分析工作流程。
避光,室温。常规运输 。请查阅批次 COA 获取详细规格。
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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。
从植物组织中制备基因组DNA较常采用的方法有氯化离心法、CTAB抽提法等。CTAB抽提法是经典的植物DNA提取法,可用于多种不同类型植物样品的DNA提取,获得的DNA产量较高,纯度虽一般,但足以满足大多数分子生物学实验需求。
CTAB 抽提液的有效成分为CTAB(十六烷基三乙基溴化铵),临用前需加入2-ME,可提升试剂有效性与稳定性。本产品仅用于科研领域,不适用于临床诊断或其他用途。
| C1518318 | Component | 500 mL | Storage |
| C1518318A | CTAB 抽提液 | 500 mL | RT. |
| C1518318B | 2-ME | 10 mL | RT. Store in the dark. |
自备材料
操作步骤(仅供参考)
1. 取5 mL适量的CTAB 抽提液,按CTAB 抽提液:2-ME=50:1的比例混匀,置于15 mL或其他规格的离心管中,60℃预热;如有必要,可加入1~5 μg/mL的RNase A,以去除残余RNA。
2. 称取1~1.5 g或适量新鲜植物组织或叶片,用预冷的液氮或干冰冷却研钵或匀浆器,将植物组织粉碎成细粉末,随后将冷冻组织转移至离心管中。
3. 向粉碎后的组织中按4~5 mL/g的比例加入预热的CTAB 抽提液,充分混匀后,65℃温育15~60 min,期间不时混匀。
4. 加入等体积的氯仿/异戊醇(24:1),颠倒离心管充分混匀,8000g离心5~10 min,回收上层水相(即上清液,含有所需DNA)。
5. 将上清液转移至新的离心管中,加入1/2~2/3体积预冷的异丙醇,轻轻混匀,室温静置使核酸沉淀至管底;若未观察到沉淀,可在室温下静置数小时至过夜。
6. 2000g离心2 min,轻轻弃去上清液。
7. 在松散的DNA沉淀物上加入75%乙醇,室温静置20 min,4000g离心10 min,轻轻弃去上清液。
8. 自然干燥DNA,加入适量去离子水或TE缓冲液溶解;如有必要,可加入1~5 μg/mL的RNase A去除残余RNA,最终于-20℃保存。
注意事项
1. 若每次使用量较少,可将试剂适当分装后使用,避免反复冻融或污染。
2. 为保障您的安全和健康,操作时请穿戴实验服并佩戴一次性手套。
3. 试剂开封后请尽快使用,以免影响后续实验效果。
Common methods for extracting genomic DNA from plant tissues include cesium chloride centrifugation, CTAB extraction, and others. The CTAB extraction method is a classic approach for plant DNA extraction and can be used for various types of plant samples. This method yields a high amount of DNA, and although the purity is generally moderate, it is sufficient for most molecular biology experiments.
The active component of the CTAB Extraction Buffer is CTAB (hexadecyltrimethylammonium bromide). Before use, 2-ME must be added to enhance the effectiveness and stability of the reagent. This product is intended for scientific research only and is not suitable for clinical diagnosis or other purposes.
| C1518318 | Component | 500 mL | Storage |
| C1518318A | CTAB Extraction Buffer | 500 mL | RT. |
| C1518318B | 2-ME | 10 mL | RT. Store in the dark. |
Materials to Prepare
Procedure (For Reference Only)
Take an appropriate amount of 5 mL of CTAB Extraction Buffer and mix it with 2-ME in a ratio of 50:1 (CTAB Extraction Buffer:2-ME). Place the mixture in a 15 mL or other appropriate centrifuge tube and preheat it to 60°C. If necessary, add 1–5 μg/mL of RNase A to remove residual RNA.
Weigh 1–1.5 g or an appropriate amount of fresh plant tissue or leaves. Use pre-chilled liquid nitrogen or dry ice to cool the mortar or homogenizer, and grind the plant tissue into a fine powder. Then, transfer the frozen tissue into the centrifuge tube.
Add the preheated CTAB Extraction Buffer to the powdered tissue at a ratio of 4–5 mL/g. Mix thoroughly and incubate at 65°C for 15–60 min, mixing occasionally during incubation.
Add an equal volume of chloroform/isoamyl alcohol (24:1). Invert the centrifuge tube to mix thoroughly. Centrifuge at 8000g for 5–10 min and recover the upper aqueous phase (supernatant containing the desired DNA).
Transfer the supernatant to a new centrifuge tube. Add 1/2 to 2/3 volumes of pre-chilled isopropanol, mix gently, and let it stand at room temperature to allow nucleic acid precipitation at the bottom of the tube. If no precipitate is observed, let it stand at room temperature for several hours or overnight.
Centrifuge at 2000g for 2 min and carefully discard the supernatant.
Add 75% ethanol to the loose DNA pellet, let it stand at room temperature for 20 min, and centrifuge at 4000g for 10 min. Carefully discard the supernatant.
Air-dry the DNA naturally and dissolve it in an appropriate amount of deionized water or TE buffer. If necessary, add 1–5 μg/mL of RNase A to remove residual RNA. Finally, store at -20°C.
Precautions
If the reagent is used in small quantities each time, it is recommended to aliquot it appropriately to avoid repeated freeze-thaw cycles or contamination.
To ensure your safety and health, wear laboratory gloves and a lab coat during operation.
Once opened, use the reagent as soon as possible to avoid compromising subsequent experimental results.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | C1518318 | |
| 分析证书 | C1518318 |
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