Thermo Scientific Phire 植物直接 PCR 试剂盒可直接从植物样品扩增 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。此款 DNA 聚合酶的独特特性使其性质极其稳定,能够耐受植物、真菌和细菌中存在的多种 PCR 抑制剂。
该试剂盒含有一整套经过优化的 PCR 试剂以及针对不同类型样品的详细实验方案,因而成为植物 DNA 扩增的理想选择。此外,该试剂盒还含有阳性对照反应的对照引物。该对照引物可扩增高度保守的叶绿体 DNA 区域,同时适用于多种植物物种。
产品亮点
• 无需耗时且昂贵的 DNA 纯化步骤 • 所需的样品材料极少 • 针对不同应用提供了两种简单的实验方案 • Phire 热启动 II DNA 聚合酶可以较短的延伸时间提供更高的特异性产物产量 (20 s/kb)。
应用
• 基因分型 • 转基因检测
Phire DNA 聚合酶的最佳退火温度可能与 Taq 类聚合酶不同。为了获得最佳结果,请使用我们的 Tm 计算器准确计算 Tm。
仅供科研使用。不可用于诊断程序。
规格
热启动内置热启动
反应次数500 次反应
突出端平末端
聚合酶Phire 热启动 II DNA 聚合酶
产品类型Direct PCR 试剂盒
数量500 reactions
反应形式分离组分
样品类型植物
运输条件湿冰或干冰
原始材料DNA
高 GC PCR 扩增效果中
PCR 方法直接 PCR
反应速度快速
Unit SizeEach
内容与储存
包含: • 200 μL Phire 热启动 II DNA 聚合酶 • 5 x 1.25 mL 2X Phire 植物组织 PCR 缓冲液(含 dNTP 和MgCl2) • 40 μL 通用型对照引物混合物(各 25 μM) • 2 x 5 mL 稀释液
-20°C 储存。
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.