Thermo Scientific Phire 植物直接 PCR 预混液设计用于直接从植物样本中扩增 DNA。在 PCR 之前无需进行 DNA 纯化。该试剂盒基于 Phire热启动 II DNA 聚合酶,一种含 DNA 结合域的特别设计的酶。该 DNA 聚合酶的独特特点使其极稳定,可耐受植物材料中含有的多种 PCR 抑制剂,如复合多糖、多酚和其他物质。
Phire 植物直接 PCR 预混液已经采用各种植物物种的叶片和种子以及真菌、细菌和酵母菌进行了测试。该试剂盒包括一整套优化的试剂和详细的实验方案,使其成为植物 DNA 扩增的理想选择。
主要特点
•直接 PCR—将样品直接加入到 PCR 反应中,因此不需要耗时且昂贵的 DNA 纯化步骤。 •可用的稀释&储存方案—允许从一个微小样品中进行数十次 PCR 反应,并允许重新测试 • 专特别设计的 Phire 热启动 II DNA 聚合酶—需要的 PCR 实验方案时间非常短 • 带有预混凝胶上样染料的预混液—将交叉污染的可能性降至最低,减少了样品处理,并允许直接进行凝胶上样
包含: • 10 x 1.25 mL Phire 植物直接 PCR 预混液(2X) •2 x 5 mL 稀释缓冲液 •40μ L 对照引物混合物 (每个25 μM ) •10 x 1.25 mL 水、不含核酸酶 •50μg 的 O’GeneRuler 表达 DNA 阶梯(足够用于100个应用)
储存在-20°C 下。
常见问题解答 (FAQ)
What is your recommendation to take a sample from plant or animal tissue for Direct PCR?
In order to obtain best results from Direct PCR, it is important to use a very small amount of starting materials. The recommended size is 0.3-0.5 mm in diameter. We recommend using Tissue Puncher 0.3 mm (Cat. No. F-200S), which is a convenient tool for sampling and is suitable for both plant and animal tissues.
Why are Control primers provided in the Direct PCR Kits?
It is very important to include a control reaction in the Direct PCR to ensure that the PCR conditions are optimal for the specific tissue or plant material used. Control primers are universal primers that amplify a small fragment from a conserved genome region. If the reaction with control primers fails, the sample material may not be suitable for Direct PCR, or PCR conditions need optimization.
Can I use plant or tissue sample which is not on the list of the Direct PCR Master Mix and Kit tested samples?
You can test any plant or tissue sample. However, it is recommended to first optimize the reaction. When working with new sample materials, start with the Dilution & Storage protocol as it allows several PCR reactions to be performed from the same sample during optimization. It is recommended to perform a positive control with purified sample DNA to ensure that the PCR conditions are working. If the positive control with purified DNA fails, the PCR conditions should be optimized before continuing further.
Can you amplify the same sample materials with the Direct PCR Master Mixes as with Direct PCR Kits?
Yes, the Direct PCR Master Mixes are upgrades of the Direct PCR kits and thus are suitable for the same sample materials.
Do the Direct PCR Master Mixes and Kits come with tissue punchers?
No, tissue punchers are not provided. You can purchase 0.3 mm Tissue Punchers (Cat. No. F-200S), or cut the sample with a scalpel. Some scientists use 100 µL pipette tips to take a sample.