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World File Search SystemLightCycler
LightCycler® 多重 DNA 主控
2. 如何使用本产品................................................................................................................................................5 2.1. 开始之前....................................................................................................................................................................5 样品材料...................................................................................................................................................................................5 对照反应...................................................................................................................................................................................5 引物...................................................................................................................................................................................................5 探针...................................................................................................................................................................................................5 Mg 2+ 浓度....................................................................................................................................................................................5 一般注意事项...................................................................................................................................................................5 注意事项...................................................................................................................................................................................5 2.2.协议................................................................................................................................................................................................................ 6 LightCycler ® PRO、LightCycler ® 480 和 LightCycler ® 96 系统协议........................................................................ 6 LightCycler ® PRO 系统（多孔板 96 或 384）的使用协议......................................................................... 6 LightCycler ® 480 仪器 II（多孔板 96 或 384）的使用协议......................................................................... 7 LightCycler ® 96 仪器的使用协议.........................................................................................................................10 LightCycler ® PRO、LightCycler ® 480 和 LightCycler ® 96 仪器的 qPCR 反应设置.............................................................................................11 LightCycler ® 2.0 仪器的使用协议.........................................................................................................................12 LightCycler ® 2.0 仪器的 qPCR 反应设置仪器.................................................................................................15
faststart Essential DNA绿色主
2。How to Use this Product.....................................................................................................................5 2.1.Before you Begin................................................................................................................................................................................... 5 Sample Materials................................................................................................................................................................................... 5 Control反应............................................................................................................................................................................................................................................................................................................................................................................................................................................................................................. Concentration.............................................................................................................................................................................. 5 Prevention of Carryover Contamination........................................................................................................................................ 5 2.2.Protocols................................................................................................................................................................................................... 6 LightCycler ® PRO and LightCycler ® 96 System protocols..................................................................................................... 6 Protocol for use with the LightCycler ® PRO System (Multiwell Plate 96 or 384).......................................................... 6 Protocol for use with the LightCycler ® 96 Instrument............................................................................................................. 7 Setup of the qPCR reaction for the LightCycler ® PRO and LightCycler ® 96 Instruments.......................................... 7 Two-Step RT-PCR.................................................................................................................................................................................. 8
humangenetische pcr检验
所有LightMix®套件都包含正跑辊。TREC KREC标准系列（RUO*）可用于新生儿面板。LightCycler®快速启动DNA Master Hybproge或LightCycler®多路复用DNA Master不包括在套件中。COBAS®因子II和因子V试剂盒包含PCR主混合物和正滚子辊。
不同葡萄品种的年轻和成熟叶片中线粒体和叶绿体DNA的相对拷贝数
摘要。亚细胞细胞器（植物）的DNA矩阵的拷贝数可以作为光合作用和氧化磷酸化过程的强度的指标。我们评估了三种葡萄品种的年轻和成熟叶子中线粒体和叶绿体DNA的相对拷贝数（RCN）：“ Traminer Pink”，“ Chardonnay”和“ Syrah”和“ Syrah”，在田间条件下生长。叶样品（5-10 mg），以进行随后的总DNA提取。使用LightCycler 480 SYBR Green I Master Mix（Lifescience，Roche）和LightCycler 96自动分析仪（Roche Life Science）进行QRT-PCR反应。使用GAPDH基因（染色体DNA）确定NAD1基因（线粒体DNA）和RPS16基因（叶绿体DNA）的相对拷贝数。使用2 -∆ CT 2- ∆ΔCT算法进行定量评估。已经确定，叶绿体和线粒体DNA的相对拷贝数（RCN）值变化，并取决于葡萄的品种和叶片成熟度。RCN在成熟的葡萄叶中的光合作用强度和成熟葡萄叶片的氧化磷酸化强度更高。在评估宏观能力平衡（MEB）指标时，可以得出结论，通过光合作用过程在叶绿体中获得的能量的2％至4％用于生产年轻叶子和成熟叶片中各种葡萄品种的线粒体中的宏观能。我们开发的实验方案可以成功用作测试系统，以评估各种葡萄品种的潜在产量。
DNA甲基化分析DPCRDNA甲基化分析DPCR
高-GC区域（例如RASSF1A启动子）的扩增通常效率低下，即使在未甲基化的区域中也是如此（请参阅下图中图中的第一组数据；测试的GC的GC含量为72％GC）。这可能会使不论甲基化状态如何准确量化模板的数量。然而，在使用DigitalLightCycler®DPCR系统时，将高GC增强子添加到扩增反应中可以显着提高放大效率和定量准确性（请参见下文）；这增加了基于DPCR的DNA甲基化分析的灵敏度和准确性。