计算溶液所需的质量、体积或浓度。
BioReagent,for DNA and RNA applications,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。
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。
双链 DNA 变性缓冲液(Double-stranded DNA Denaturation Buffer,常用简写:dsDNA Denaturation Buffer),又称中性转移缓冲液,主要由 1.5 M NaCl、微量 NaOH 等成分组成,不含蔗糖。本试剂专为核酸杂交实验设计,尤其适用于Southern blot实验中双链 DNA 的变性。
本产品仅用于科研领域,不适用于临床诊断或其他用途。
操作步骤(仅供参考)
可根据实验具体需求直接操作;以下为针对不同膜类型的 DNA 变性及转移专用步骤:
1. 转移至不带电荷的膜
1.1 DNA 变性:将凝胶置于10 倍凝胶体积的双链 DNA 变性缓冲液中,室温放置45 min,期间持续轻轻振荡。
1.2 中和处理:用去离子水短暂浸泡凝胶;随后将凝胶浸没于10 倍凝胶体积的 DNA 中和缓冲液 Ⅰ 中,室温放置30 min,轻轻振荡;更换一次新鲜的 DNA 中和缓冲液 Ⅰ,继续浸泡15 min。
2. 转移至带电荷的尼龙膜
2.1 碱性变性:将凝胶置于5-10 倍凝胶体积的 DNA 碱性转移缓冲液中,室温放置15 min,持续轻轻振荡。
2.2 二次变性:更换一次新鲜的 DNA 碱性转移缓冲液,继续浸泡20 min,期间不断轻轻振荡。
注意事项
1. 若目标 DNA 片段长度小于 20 kb,建议避免进行脱嘌呤反应,以防片段降解。
2. 单次使用量较小时,建议将试剂适当分装后使用,减少反复冻融对试剂稳定性的影响。
3. 试剂开封后请尽快使用,避免试剂失效或污染,影响后续核酸转移及杂交实验效果。
Double-stranded DNA Denaturation Buffer, also known as Neutral Transfer Buffer, is primarily composed of 1.5 M NaCl, trace NaOH, and other components, and does not contain sucrose. This reagent is specifically designed for nucleic acid hybridization experiments, particularly suitable for the denaturation of double-stranded DNA in Southern blot experiments.
This product is for research use only. Not for use in diagnostic procedures or other purposes.
Procedure (For Reference Only)
The product can be used directly according to specific experimental requirements. The following are dedicated steps for DNA denaturation and transfer for different membrane types:
1. Transfer to Uncharged Membranes
1.1 DNA Denaturation: Place the gel in 10 volumes of Double-stranded DNA Denaturation Buffer. Incubate at room temperature for 45 minutes with continuous gentle agitation.
1.2 Neutralization Treatment: Briefly rinse the gel with deionized water. Subsequently, submerge the gel in 10 volumes of DNA Neutralization Buffer I. Incubate at room temperature for 30 minutes with gentle agitation. Replace with fresh DNA Neutralization Buffer I and continue incubation for another 15 minutes.
2. Transfer to Charged Nylon Membranes
2.1 Alkaline Denaturation: Place the gel in 5-10 volumes of DNA Alkaline Transfer Buffer. Incubate at room temperature for 15 minutes with continuous gentle agitation.
2.2 Secondary Denaturation: Replace with fresh DNA Alkaline Transfer Buffer and continue incubation for another 20 minutes with gentle agitation.
Notes
If the target DNA fragment length is less than 20 kb, it is recommended to avoid performing depurination reactions to prevent fragment degradation.
If the single-use volume is small, it is recommended to aliquot the reagent appropriately before use to minimize the impact of repeated freezing and thawing on reagent stability.
Use the reagent as soon as possible after opening to avoid reagent inactivation or contamination, which could affect subsequent nucleic acid transfer and hybridization experiment results.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | D1518770 |
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