BioReagent,分子生物学级,即用型,用于核酸电泳,用于聚丙烯酰胺凝胶电泳 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
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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。
DNA Marker由纯化的不同分子量双链DNA组成，为即用型分子量标准品。通过电泳后条带的位置和亮度，可预估样品的分子量大小及浓度。本产品已预混上样缓冲液，可直接上样电泳。本产品除了琼脂糖电泳验证外，大多数条带都经过严格的PAGE检验，PAGE胶中无“双眼皮”，同一大小的DNA分子跑出多条带等现象。当琼脂糖凝胶中含大分子染料（如Gelred）时，加入电泳增强剂（主要成分为大分子核酸染料）能有效减少Marker在电泳时的拖带现象，也可选购阿拉丁Gelred-prestained 系列产品 。
使用方法：
取5μL Marker加入琼脂糖凝胶加样孔（5mm）进行电泳；上样量可根据孔径调整。
注意事项：
“拖带现象”是琼脂糖凝胶中加入大分子染料（如GelRed）电泳时的一个常见现象，经常表现为小分子量条带相对较亮，大分子量条带相对较暗，整体条带下坠，条带间粘连等。我们的实验工作揭示造成此现象的本质原因是凝胶中的染料相对不足。
与EB等小分子量水溶性染料不同，GelRed在水中的溶解度极低，市售推荐的染料使用浓度跟EB的推荐使用浓度相差较大。当含GelRed的凝胶第一次被使用时，一般还是可以获得比较好的电泳效果，一旦凝胶被重复使用或浸泡在电泳液中时间过久，凝胶中的染料被释放至电泳液中，导致胶中染料浓度进一步降低，小分子DNA条带先进入凝胶结合染料后，后面大分子DNA再进入凝胶中时，供结合的染料相对不足，造成“拖带现象”。
改善琼脂糖凝胶 (含大分子核酸染料) 电泳时 “拖带现象” 的两个方向
| 增加染料相对浓度 | 减少染料损失 |
| ⁕提前在Marker中加入电泳增强剂 (主要成分是核酸染料)。 ⁕凝胶中可增加染料浓度，如10000x染料按5000x使用。 ⁕减少DNA Marker的上样量。 | ⁕凝胶不要重复使用多次 (电泳时染料向上迁移出凝胶)。 ⁕凝胶不要长时间浸泡在电泳液中，导致染料析出。 |
DNA Marker consists of purified double-stranded DNA of various molecular weights and is a ready-to-use molecular weight standard. The molecular weight and concentration of samples can be estimated based on the position and brightness of bands after electrophoresis. This product is pre-mixed with loading buffer and can be directly loaded for electrophoresis. In addition to verification by agarose electrophoresis, most bands have undergone strict PAGE testing, with no issues such as "double bands" or multiple bands for DNA molecules of the same size observed in PAGE gels. When agarose gels contain macromolecular dyes (e.g., GelRed), adding an electrophoresis enhancer (main component: macromolecular nucleic acid dye) can effectively reduce band smearing of the Marker during electrophoresis. Aladdin GelRed-prestained series products are also available for selection.
Experimental procedures:
Add 5 μL of Marker to the agarose gel well (5 mm) for electrophoresis; the loading volume can be adjusted according to the well size.
Precautions:
"Band smearing" is a common phenomenon during electrophoresis when macromolecular dyes (e.g., GelRed) are added to agarose gels. It often manifests as relatively bright low-molecular-weight bands, relatively dim high-molecular-weight bands, overall band sagging, and adhesion between bands. Our experiments reveal that the root cause of this phenomenon is insufficient dye concentration in the gel.
Unlike small-molecular-weight water-soluble dyes such as EB, GelRed has extremely low solubility in water, and the recommended working concentration of commercially available GelRed differs greatly from that of EB. Good electrophoresis results are generally obtained when a GelRed-containing gel is first used. However, if the gel is reused or soaked in electrophoresis buffer for too long, the dye in the gel leaches into the buffer, further reducing the dye concentration in the gel. Low-molecular-weight DNA bands enter the gel first and bind to the dye, leaving insufficient dye for subsequent high-molecular-weight DNA, resulting in "band smearing".
Two Approaches to Improve "Band Smearing" in Agarose Gel Electrophoresis (Containing Macromolecular Nucleic Acid Dyes)
| Increase the relative dye concentration | Reduce dye loss |
| ⁕ Add electrophoresis enhancer (main component: nucleic acid dye) to the Marker in advance. ⁕ Increase the dye concentration in the gel (e.g., use 10000× dye at 5000×). ⁕ Reduce the loading volume of DNA Marker. | ⁕ Avoid repeated use of the gel (dye migrates upward out of the gel during electrophoresis). ⁕ Prevent prolonged soaking of the gel in electrophoresis buffer to avoid dye leaching. |
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0635994 |
分析证书 | D1521306 |
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 Ready-to-use 级别指南 → 查看 Suitable for molecular biology 级别指南 → 查看 for NA electrophoresis 级别指南 → 查看 for PAGE 级别指南 →
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