| The LavaLAMP™ DNA Master Mix is intended to simplify development and optimization of DNA LAMP (loop-mediated isothermal amplification) reactions. LAMP kits are commonly available as multicomponent kits that require optimization (e.g. MgSO4, betaine, enzyme as well as temperature, primer concentration, etc.). The LavaLAMP™ DNA Master Mix greatly simplifies reaction optimization by limiting optimization to target specific components/conditions (LAMP primers, target concentration and reaction temperature). Additionally, this master mix is heat stable at 90˚C for ≤5 minutes, which enables the addition of a reaction preheating step to help reduce the time to result (TTR) when using purified target DNA as input. This kit is for research purposes only, and available under the limited-use license described at the end of this document.LAMP commonly employs a set of six primers (F3, B3, Loop-F, Loop-B, FIP and BIP), which must be supplied by the user. Lucigen recommends using previously-established designs or designing new primer sets using the Eiken web utility (see Appendix A). Not all primer sets identified by this program are guaranteed to perform with LavaLAMP™ DNA Master Mix or any other LAMP system. We recommended that you design multiple primer sets to identify the best performing set. We also highly recommend inclusion of loop primers (Nagamine, 2002). LavaLAMP™ DNA Master Mix amplified products may be detected by agarose gel electrophoresis or by real-time/end-point monitoring using fluorescent double-stranded DNA-binding dyes, such as the Green Fluorescent Dye included in Cat. No. 30067-1. Turbidity may also be used (Mori, 2001), but this method is less sensitive. LavaLAMP™DNA预混液旨在简化DNA LAMP（环介导的等温扩增）反应的开发和优化。 LAMP试剂盒通常是需要优化的多组分试剂盒（例如MgSO4，甜菜碱，酶以及温度，引物浓度等）。 LavaLAMP™DNA预混液通过将优化限制在特定目标组分/条件（LAMP引物，目标浓度和反应温度）上，大大简化了反应优化过程。 此外，该预混合物在90°C的温度下热稳定≤5分钟，这使得添加反应预热步骤有助于减少使用纯化的靶DNA作为输入时的生成时间（TTR）。 该套件仅用于研究目的，可根据本文档末尾所述的有限使用许可使用。 LAMP通常使用一组六个引物（F3，B3，Loop-F，Loop-B，FIP和BIP），这些引物必须由用户提供。 Lucigen建议使用以前建立的设计或使用Eiken Web实用程序设计新的引物组（请参阅附录A）。 不能保证该程序确定的所有引物都可与LavaLAMP™DNA Master Mix或任何其他LAMP系统一起使用。 我们建议您设计多个引物组以鉴定最佳表现组。 我们还强烈建议包括环状引物（Nagamine，2002）。 LavaLAMP™DNA Master Mix扩增产物可以通过琼脂糖凝胶电泳或通过使用荧光双链DNA结合染料（例如货号 30067-1中包含的Green Fluorescent Dye）的实时/终点监测来检测。 也可以使用浊度（Mori，2001），但是这种方法不那么敏感。 |
| 优点： |
| ▪ Loop-mediated isothermal amplification (LAMP): Facilitates running amplification reactions in difficult test environments, enabling use outside of a high-tech laboratory.▪ Master Mix Format: Simplifies reaction setup and minimizes optimization required to produce the best assay results.▪ High Thermal Stability: Highly thermostable enzyme (up to 90°C for 5 min) enables preheating of reactions containing purified target DNA which,depending on the target, may increase assay sensitivity and decrease time to results.▪ Elevated LAMP Reaction Temperature (68-74°C): Improves primer specificity and reduces background amplification depending on the DNA target.▪ Freeze-dry Compatible: Enables generation of room temperature stable test kits through lyophilization. ▪ 环-介导的等温扩增（LAMP）：便于在困难的测试环境中进行扩增反应，从而可以在高科技实验室之外使用。 ▪ 预混液格式：简化反应设置，并最小化产生最佳测定结果所需的优化。 ▪ 较高的热稳定性：高度耐热的酶（在90°C下持续5分钟）可预热含有纯化目标DNA的反应，具体取决于目标DNA，可增加测定的灵敏度并缩短获得结果的时间。 ▪ 升高的LAMP反应温度（68-74°C）：根据DNA靶点，提高引物特异性并减少背景扩增。 ▪ 兼容冻干：通过冻干可生成室温稳定的测试试剂盒。 |
| 数据： |
| Figure 1: Positive LAMP Reaction Products from a Positive Control Target. Lane 1: 100 bp Marker, Lane 2: No Target Control reaction. Lanes 3 and 4: A distinct banding pattern is seen among the smear, which is indicative of a positive LAMP reaction.图1：来自阳性对照靶的阳性LAMP反应产物。 泳道1：100bp标记，泳道2：无靶对照反应。 泳道3和4：在涂片中看到明显的条带图案，这表明LAMP反应为阳性。Figure 2: Typical Background Amplification in a LAMP Reaction. Lane 1: 100 bp ladder, Lanes 2 and 3: Non-specific or background amplification appears as a smear of DNA fragments with no visible or distinct bands. A prominent primer dimer band is also characteristic of non-specific amplification. Lane 4: Absence of non-specific amplification (no products).图2：LAMP反应中的典型背景扩增。 泳道1：100 bp梯形图，泳道2和3：非特异性或背景扩增表现为DNA片段涂片，无可见或明显的条带。 突出的引物二聚体带也是非特异性扩增的特征。 泳道4：没有非特异性扩增（无产物）。 |
| 组成成分： |
| ▪ LavaLAMP™ DNA Master Mix：-20°C▪ DNA Positive Control LAMP Primer Mix：-20°C▪ DNA Positive Control：-20°C |
| 相关产品： |
| ▪LavaLAMP™ DNA Component Kit（#30076）▪Green Fluorescent Dye（#30078） ▪ LavaLAMP™ DNA Master Mix with Dye(#30067-1) |
| 参考文献： |
| 1.Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res. 2000 28(12):E63. 2.Nagamine K, Hase T, Notomi T. Accelerated reaction by loop-mediated isothermal amplification using loop primers. Mol Cell Probes. 2002 16(3):223-9. 3.Mori Y, Nagamine K, Tomita N, Notomi T. Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem Biophys Res Commun. 2001 289(1):150-4. 4.Mori Y, Hirano T, Notomi T. Sequence specific visual detection of LAMP reactions by addition of cationic polymers. BMC Biotechnol. 2006 6:3. 5.Tomita N, Mori Y, Kanda H, Notomi T. Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products. Nat Protoc. 2008 3(5):877-82. |
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