DuFinder是一种新型核酸染料，这种独特的油性大分子，不易挥发升华、不易吸入人体，不能穿透细胞膜进入活体细胞内，且在凝胶染色浓度下没有诱变性，具有使用安全、检测灵敏等特点，可以作为各种核酸电泳的染色剂，适用于各种片段大小染色。与标准凝胶成像系统和可见光激发的凝胶观察装置完美兼容，适用于紫外凝胶成像系统或蓝色可见光激发的凝胶观察装置。本公司提供的DuFinder荧光染料为浓缩的10000×染料。
操作步骤：
1. 琼脂糖凝胶电泳染色 (推荐方法)
将 DuFinder核酸染料 (10000×) 加入凝
① 制胶：按常规操作，制备琼脂糖凝胶，加入浓缩的 DuFinder核酸染料 (10000×)，使其在凝胶中的终浓度为1×。
(例如：制备50ml的凝胶，加入染料5μl)，轻轻摇匀，倒胶。
② 按常规方法电泳，观测结果 (染料会使DNA迁移变慢，所以可适当加大电压进行电泳)。
2. 泡染法
① 按照常规方法进行电泳。
② 用H₂O将 DuFinder核酸染料 (10000×) 稀释约3300倍到0.1M 的NaCI中，制成3×染色液。
(例如将15μL DuFinder核酸染料 (10000×) 和5ml 1M NaC加到 45ml H₂O中)。
③ 将凝胶小心地放入合适的容器中，如聚丙烯容器中。缓慢加入足量的3×染色液浸没胶。室温振荡染色30min左右。
④ 在凝胶成像仪内，观测结果。
注意事项：
请务必在使用本试剂盒之前阅读此注意事项！
1. 由于DuFinder 具有良好的热稳定性，可以在热的琼脂糖溶液中直接添加，而不需要等待溶液冷却。摇晃，振荡或者翻转以保证染料充分混匀。也可以选择将DuFinder储液加到琼脂糖粉末和电泳缓冲液中，然后用微波炉或其他常用方式加热以制备琼脂糖凝胶。DuFinder 兼容所有常用的电泳缓冲溶液。
2. 如果总是看到条带弥散或分离不理想，建议使用泡染法染色以确认问题是否与染料有关。如果染色后问题依旧存在，则说明问题与染料无关，请尝试：降低琼脂糖浓度；选用更长的凝胶；延长凝胶时间以保证边缘清晰；改进上样技巧或选择泡染法染色。
3. DuFinder 对玻璃器皿和非聚丙烯材料具有一定的亲合力。建议在稀释、贮存、染色等使用过程中用聚丙烯类容器。
4. 对于聚丙烯酰胺凝胶请使用泡染法。
DuFinder is a novel nucleic acid stain. As a unique oily macromolecule, it features low volatility and sublimation, low risk of inhalation into the human body, and cannot cross cell membranes to enter living cells. At gel staining concentrations, it is non-mutagenic. Characterized by safe operation and high detection sensitivity, it can be used as a stain for various nucleic acid electrophoresis and is suitable for staining nucleic acid fragments of different sizes. It is fully compatible with standard gel imaging systems and visible-light-excited gel viewers, applicable to both UV gel imaging systems and blue visible light-excited gel observation devices. The DuFinder fluorescent stain supplied by our company is a concentrated 10000× dye.
Operating Procedures:
1. Agarose Gel Electrophoresis Staining (Recommended Method)
① Gel casting: Prepare agarose gel following standard protocols. Add concentrated DuFinder nucleic acid dye (10000×) to reach a final concentration of 1× in the gel. (Example: Add 5 μL of dye for 50 mL gel solution). Mix gently and cast the gel.
② Perform electrophoresis routinely and visualize results. (The dye slows DNA migration; the voltage may be increased appropriately during electrophoresis.)
2. Post-electrophoresis Bath Staining
① Run electrophoresis using standard protocols.
② DuFinder nucleic acid dye (10000×) approximately 3300-fold in 0.1 M NaCl solution to prepare 3× staining solution. (Example: Add 15 μL DuFinder nucleic acid dye (10000×) and 5 mL 1 M NaCl into 45 mL H₂O.)
③ Carefully place the gel into an appropriate container such as a polypropylene vessel. Slowly add sufficient 3× staining solution to fully submerge the gel. Shake at room temperature for approximately 30 min for staining.
④ Visualize bands using a gel imager.
Precautions:
Please read these notes thoroughly before using this kit!
1. DuFinder has excellent thermal stability. It can be added directly into hot agarose solution without waiting for cooling. Shake, vortex or invert the solution to ensure thorough mixing. Alternatively, DuFinder stock solution may be added to agarose powder and electrophoresis buffer before heating by microwave or other common methods for gel preparation. DuFinder is compatible with all commonly used electrophoresis buffers.
2. If band smearing or poor separation persists, bath staining is recommended to verify whether the issue originates from the dye. If the problem remains after staining, it is unrelated to the dye. Troubleshooting options: reduce agarose concentration; use a longer gel; extend gelation time for sharp band edges; optimize sample loading technique, or adopt bath staining.
3. DuFinder has certain affinity for glassware and non-polypropylene materials. Polypropylene containers are recommended for dilution, storage and staining operations.
4. Use bath staining for polyacrylamide gels.
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