计算溶液所需的质量、体积或浓度。
BioReagent,分子生物学级,无菌 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,-20°C储存。超低温运输 。请查阅批次 COA 获取详细规格。
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。
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。
DNA 寡核苷酸退火缓冲液(Annealing Buffer for DNA Oligos,简称 DNA 退火缓冲液),适用于常规 DNA oligo 退火,尤其适合RNAi/siRNA 质粒构建中较难退火的 DNA oligo。此类 DNA oligo 因常含约 20 bp 互补序列,易自身形成发夹结构,影响两条链正确配对退火。
DNA 寡核苷酸退火缓冲液(5×)可有效抑制 DNA oligo 自身发夹结构形成,退火产物可直接用于酶切后质粒的连接与转化,能显著提高阳性克隆得率。本试剂操作简便,只需将 oligo 与退火缓冲液按比例混合,经 PCR 仪程序降温约 90 min 即可完成退火。
本产品仅用于科研领域,不适用于临床诊断或其他用途。
| D1518364 | Component | 1 mL | Storage |
| D1518364A | DNA寡核苷酸退火缓冲液 (5×) | 1 mL | -20℃. |
| D1518364B | 无核酸酶水 | 5 mL | RT. |
操作步骤(仅供参考)
1. 试剂准备
将待退火的 DNA oligo 用无菌超纯水或无核酸酶水配制成 50 μM 工作液;将 Annealing Buffer for DNA Oligos (5×) 溶解并充分混匀备用。
2. 按如下退火反应体系操作(总体积 100 μL)
|
按上述顺序依次加样并混匀;若 PCR 仪无热盖,可滴加Mineral Oil防止蒸发。
3. PCR 仪退火程序
|
退火产物可直接用于连接反应,或 -20℃ 冻存备用。若需进行酶切等后续酶反应,建议先纯化退火产物,或保证退火产物在体系中体积占比<5%,避免缓冲液干扰酶活。
注意事项
1. 试剂避免反复冻融,以防退火效率下降。
2. 本产品仅适用于 DNA oligo 退火,不可用于 RNA oligo。
3. 操作时请穿戴实验服与一次性手套,保证实验安全。
4. 试剂开封后建议尽快使用,以免影响实验效果。
Annealing Buffer for DNA Oligos is suitable for routine DNA oligo annealing, and is particularly effective for difficult-to-anneal DNA oligos used in RNAi/siRNA plasmid construction. Such DNA oligos often contain approximately 20 bp complementary sequences, making them prone to forming hairpin structures that can interfere with correct pairing and annealing of the two strands.
Annealing Buffer for DNA Oligos (5×) effectively inhibits the formation of DNA oligo hairpin structures. The annealed products can be used directly in subsequent steps such as restriction enzyme digestion followed by plasmid ligation and transformation, significantly improving the positive clone yield. This reagent is easy to use; simply mix the oligos with the annealing buffer in the appropriate proportions and complete the annealing process via a PCR instrument programmed for controlled cooling over approximately 90 minutes.
This product is for research use only. Not for use in diagnostic procedures or other purposes.
| D1518364 | Component | 1 mL | Storage |
| D1518364A | Annealing Buffer for DNA Oligos (5×) | 1 mL | -20℃. |
| D1518364B | Nuclease-Free Water | 5 mL | RT. |
Procedure (For Reference Only)
1. Reagent Preparation
Prepare the DNA oligos to be annealed as a 50 μM working solution using sterile ultrapure water or Nuclease-Free Water.
Thaw and thoroughly mix the Annealing Buffer for DNA Oligos (5×).
2. Annealing Reaction Setup (Total Volume 100 μL)
Prepare the reaction mixture according to the table below, adding components in the order listed and mixing well. If the PCR instrument does not have a heated lid, overlay with Mineral Oil to prevent evaporation.
|
3. PCR Instrument Annealing Program
|
4. Subsequent Use
The annealed product can be used directly in ligation reactions or stored at -20℃ for later use.
If subsequent enzymatic reactions (e.g., restriction enzyme digestion) are required, it is recommended to purify the annealed product first, or ensure that the volume of the annealed product constitutes less than 5% of the final reaction system to avoid buffer interference with enzyme activity.
Notes
Avoid repeated freezing and thawing of the reagent to prevent decreased annealing efficiency.
This product is specifically designed for DNA oligo annealing and should not be used for RNA oligos.
Please wear laboratory appropriate attire and disposable gloves during operation to ensure safety.
Use the reagent as soon as possible after opening to ensure optimal performance.
通过匹配包装上的批号来查找并下载产品的 COA,每批产品都进行了严格的验证,您可放心使用!
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | D1518364 |
我们的级别选择指南涵盖了目录中所有的变体的纯度、稳定剂状态和应用适用性。
查看 BioReagent 级别指南 → 查看 Suitable for molecular biology 级别指南 → 查看 Sterile 级别指南 →