The protocol was not followed correctly Verify that all steps have been followed and the correct reagents, dilutions, volumes, and cycling parameters have been used. Template contains inhibitors, nucleases, or
The buffer is also optimized for increased resistance to reaction inhibitors originating from sample material or nucleic acid purification steps. Online resources thermofisher.com/lyo-ready For further information, contact MDxenzymes@thermofisher.com Protocol Please follow the instructions below to prepare ...
The protocol was not followed correctly Verify that all steps have been followed and the correct reagents, dilutions, volumes, and cycling parameters have been used. Template contains inhibitors, nucleases, or proteases, or has otherwise been degraded. Pu...
Design primers and probes Check primer specificity Assess primer and probe properties: melting temperature (Tm), secondary structure, and complementarity Determine PCR product properties Validate the primers and/or probes and optimize the protocol ...
When you want rapid and cost-effective results in a single-tube protocol, utilize our One-Step TB Green PrimeScript RT-PCR Kit II (Perfect Real Time). This kit is available in both 100 and 500 reaction sizes. This method detects fluorescence produced during the amplification process by adding...
PCR反应的扩增效率严重的影响到PCR的结果,同样在qRT-PCR中,扩增效率对定量的结果尤为重要,除去反应液(reaction buffer)中其他物质及机器和protocol带来的影响,引物的质量对于qRT-PCR的扩增效率影响也非常大,为了保证结果的准确性,无论是相对荧光定量还是绝对荧光定量均需要对引物的扩增效率进行检测,而公认的有效的qRT-...
before explaining qPCR data analysis, we need to quickly discuss how to determine reaction efficiency and specificity. You don't need to perform these steps after every qPCR experiment, but should always validate these two values when setting up a new qPCR protocol or changing your current workflo...
Additionally, while the ability to optimize both the RT and qPCR steps separately can improve sensitivity and efficiency, it comes with the drawback of having to optimize two reactions instead of one. Lastly, the two-tube protocol cannot be as easily adapted to automated, high-throughput ...
France) in a 5% CO2humidified incubator at 37 °C. LCLs were seeded at 4 × 105cells/ml. After 4 days cells were harvested for RNA isolation. Total RNA was extracted from 5 × 106cells pellets using the miRNeasy Mini Kit according to the manufacturer’s protocol (QIAGEN, Fran...
PrimeTime One-Step RT-qPCR Master Mix protocol(391 KB) Brochures and flyers pdf PrimeTime One-Step RT-qPCR Master Mix Product Brochure(481 KB) Handbooks Real-Time PCR handbook Safety data sheets Search for MSDS by product name or part number ...