Can TaqMan®gene expression assays be used in duplex real-time PCR?
Duplex reactions using TaqMan®Gene Expression Assays Duplex real-time PCR is the simultaneous amplification and measurement of two target sequences in one reaction. TaqMan®Gene Expression Assays can be used in duplex real-time PCR when using a FAM™dye-labeled assay in combination with a primer-limited, VIC®dye-labeled assay.
What should I know before using TaqMan® PCR core reagents kit?
Check the settings of the dye components before data analysis. Decrease in ROX™ dye fluorescence (passive reference dye). Precipitation in the TaqMan® buffers occurs. • When using the TaqMan® PCR Core Reagents Kit, be sure to mix the tubes well. • Use TaqMan® Gene Expression Master Mix (2 ?).
What cDNA template should be used for qPCR?
10 50 cDNA Template (1-100ng)§ § Applied Biosystems recommends that the qPCR reaction not be composed of more than 20% of the reverse transcription reaction. 420 RNAse-free water 4 20 50TaqMan®Gene Expression Assays Protocol
What size amplicon should be used in qPCR reaction?
The size of amplicon in qPCR reaction is between 75-200bp to ensure higher amplification efficiency. For primer and probe design in this purpose, the software Primer Express v2.0 from ABI is highly recommended, especially for TaqMan chemistry, which needs both sequence specific primers and fluorescence labeled probe.
Estandarización de una Técnica de PCR en Tiempo Real con