Technically Speaking: Promega RT-PCR Systems ExplainedHanan Abramovici
To further understand the adaptability of our assay in diagnostic and hospital laboratories, we validated the method in three additional commonly used RT-PCR instruments with melt capability. The performance of these new test instruments was compared to the LC480 as the gold standard. A total of ...
To test our platform’s capability to perform multiplex single-cell RT-PCR on living cells encapsulated in microdroplets, three murine cell lines with various gene expression profiles, namely B62c, D4T, and EL4, were assayed. No issues were observed when encapsulating the cell lines despite the ...
Real-time PCR has been a mainstay in diagnostics over the years due to its speed, specificity, sensitivity, reproducibility and throughput; as such, real-time PCR is a target for improvement. Nevertheless, to validate a novel diagnostic tool, correct setup of the assay, including proper ...
This strategy eliminates variation introduced by pre-PCR and PCR processing. However, it has been shown that no RG is truly stably expressed over all kinds of culture conditions or among different cell types. This might be explained by the fact that housekeeping genes also participate in other ...
Real-time RT-PCR normalisation; strategies and considerations. Genes Immun. 6, 279–284 (2005). 6. Gutierrez, L. et al. The lack of a systematic validation of reference genes: a serious pitfall undervalued in reverse transcription- polymerase chain reaction (RT-PCR) analysis in plants. Plant...
A more stringent test of PCR specificity was performed by sequencing the amplification products of each of the 42 candidate reference genes. Always the sequence of the PCR product matched that of the target cDNA, thereby confirming the exquisite PCR specificity of the developed primer pairs. Figure...
For the second case, inhibition during RT-PCR reaction or difference in the melting temperature used by the two laboratories could explain the result. For this last sample, the melting curve was also significantly different from those usually expected for PoSaV. This could be explained by the ...
RT-qPCR data for 18080 protein-coding genes were generated in the context of the Sequencing Quality Control study (SEQC) (17) using PrimePCR assays (BioRad) (Supplemental Table 1). In order to define the ensemble of transcripts amplified by every individual qPCR assay, assays were re-mapped...
Reticulons (RTNs) are a large family of membrane associated proteins with various functions. NOGO-A/RTN4A has a well-known function in limiting neurite outgrowth and restricting the plasticity of the mammalian central nervous system. On the other hand, R