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A multiplex reaction for the recombination band, and the control gene GAPDH was performed for each sample.
This shows the relative simplicity of this approach to optimizing the multiplex reaction for specific needs, whether geographical or for specific disease syndromes.
cDNA for UCK2 and ACTB were amplified in a multiplex reaction for the purpose of increasing the quantity of the target cDNA prior to quantitative real-time PCR.
The amplification was performed as a multiplex reaction for SARS-CoV gRNA, sgRNA, and G3PDH by using a OneStep RT-PCR procedure (Qiagen, Inc., Valencia, CA); thus, both positive- and negative-sense SARS-CoV RNAs served as templates for reverse transcription.
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The SNP genotyping method used the Mass Array system to design multiplex reactions for PCR and iPlex primer extension (Sequenom) and the MALDI-TOF based Mass Array genotype platform (Sequenom).
We propose eight markers to combine in two multiplex reactions for further studies on a larger set of populations.
Sequenom Typer 4.0 (Sequenom, Inc., San Diego, CA, USA) was used to design four multiplex reactions for 30 SNPs within the 10 selected genes.
While this assay was run separately, it would be possible to consider including this control in the multiplex reaction by, for example, labelling both subgenotypes of BPI3 with the same fluorophore.
Commercially available primers and a VIC-TAMRA-labelled probe for the housekeeping gene RNase P were included in the multiplex reaction to standardise for DNA input.
The sensitivity was demonstrated to be 2.5 (for multiplex reaction detecting viruses from species A, C, F) and 24 (for multiplex reaction detecting viruses from species B, D, E) copies of viral genome/reaction.
One multiplex reaction was carried out for PLAT, MMP13, PAI, MMP9 and MMP2, and the other included MMP3, MMP7 and MMP1.
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