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核酸提取方法,本产品经 RNase free 处理,主要用于植物组织总 RNA 的提取,所获核酸可满足多数分子生物学实验需求。
CTAB 抽提液 (RNase free) 的有效成分为CTAB(十六烷基三乙基溴化铵),经RNase free处理,临用前加入2-ME,可提升试剂有效性与稳定性。该试剂仅用于科研领域,不适用于临床诊断或其他用途。
| C1518317 | Component | 500 mL | Storage |
| C1518317A | CTAB 抽提液 (RNase free) | 500 mL | RT. |
| C1518317B | 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或适量的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DNase,以便去除样品中的DNA。
2. 称取1.0~1.5 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,将新鲜植物组织或叶片粉碎成细粉末,将冷冻的组织转移至离心管中。
3. 向粉碎后的组织中按4~5 mL/g加入预热的CTAB抽提液,充分混匀,65℃温育15~60 min,并不时混匀。
4. 加入等体积氯仿/异戊醇 (24:1),颠倒充分混匀,8000 g离心5~10 min,回收上层水相(即上清液,该上清液中含有所需DNA/RNA)。
5. 转移上清液至新的离心管中,加入1/2~2/3体积预冷的异丙醇,轻轻混匀,室温静置使核酸沉至管底;如果观察不到沉淀,可在室温下静置数小时至过夜。
6. 2000 g离心2 min,轻轻弃上清液。
7. 在松散的DNA/RNA沉淀物上加入75%乙醇,室温静置20 min,4000 g离心10 min,轻轻弃上清液。
8. 自然干燥RNA,加入适量去离子水或TE缓冲液;如有必要可再加入1~5 μg/mL的DNase Ⅰ,以便去除残余的DNA,-20℃保存。
注意事项
1. 如果每次的使用量很小,可以适当分装后再使用。
2. 为了您的安全和健康,请穿实验服并戴一次性手套操作。
3. 试剂开封后请尽快使用,以防影响后续实验效果。
Common methods for extracting genomic DNA from plant tissues include cesium chloride centrifugation and CTAB extraction. The CTAB extraction method is a classic technique for plant nucleic acid extraction. This product is RNase-free and primarily used for total RNA extraction from plant tissues. The nucleic acids obtained can meet the requirements of most molecular biology experiments.
The active component of CTAB Extraction Buffer (RNase-free) is CTAB (cetyltrimethylammonium bromide). It is treated to be RNase-free, and 2-ME should be added before use to enhance the reagent's effectiveness and stability. This reagent is intended for research purposes only and is not suitable for clinical diagnosis or other applications.
| C1518317 | Component | 500 mL | Storage |
| C1518317A | CTAB Extraction Buffer (RNase-free) | 500 mL | RT. |
| C1518317B | 2-ME | 10 mL | RT. Store in the dark. |
Materials to Prepare
Procedure (For Reference Only)
Take 5 mL or an appropriate amount of CTAB Extraction Buffer and mix it with 2-ME at a ratio of 50:1 (CTAB Extraction Buffer:2-ME). Place the mixture in a 15 mL or other suitable centrifuge tube and preheat at 60°C. If necessary, add 1–5 μg/mL of DNase to remove DNA from the sample.
Weigh 1.0–1.5 g or an appropriate amount of fresh plant tissue or leaves. Pre-cool the mortar or homogenizer with liquid nitrogen or dry ice, and grind the fresh plant tissue or leaves into a fine powder. Transfer the frozen tissue into a centrifuge tube.
Add 4–5 mL/g of preheated CTAB Extraction Buffer to the powdered tissue, mix thoroughly, and incubate at 65°C for 15–60 minutes, mixing occasionally.
Add an equal volume of chloroform/isoamyl alcohol (24:1), mix thoroughly by inverting, and centrifuge at 8000 g for 5–10 minutes. Recover the upper aqueous phase (supernatant), which contains the desired DNA/RNA.
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 acids to settle at the bottom of the tube. If no precipitate is observed, let it stand at room temperature for several hours or overnight.
Centrifuge at 2000 g for 2 minutes, and carefully discard the supernatant.
Add 75% ethanol to the loose DNA/RNA pellet, let it stand at room temperature for 20 minutes, and centrifuge at 4000 g for 10 minutes. Carefully discard the supernatant.
Air-dry the RNA naturally and dissolve it in an appropriate amount of deionized water or TE buffer. If necessary, add 1–5 μg/mL of DNase I to remove residual DNA. Store at -20°C.
Notes
If the usage amount is small each time, the reagent can be aliquoted before use.
For your safety and health, please wear a lab coat and disposable gloves during operation.
Use the reagent as soon as possible after opening to avoid affecting subsequent experimental results.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | C1518317 |
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