Integrated DNA Technologies
Prime Time Gene Expression Master Mix
1055770
I have used Integrated DNA technologies Prime Time Gene expression master mix in a multiplexed qPCR assay to measure expression of key genes induced in Salmonella enterica under acidic stress conditions. Got consist results with higher expression of acid induction system. Prime Time Gene expression master mix comes with ready-to-use mix containing all the components of qPCR reaction except the primers. Just need to prepare master mix using primers which you want to study. Optimal performance compared to other kits and minimizes hands on time in preparing master mix and ideal for multiplexed assays.
Multiplexed qPCR assay for gene expression in Salmonella enterica
cDNA prepared from S. enterica total RNA
Prepared master mix multiplexed qPCR reaction targeting 3 genes according the instructions given the protocol for 20 microliter reaction. Master mix can be used at room temperature. Then performed qPCR reaction in agilent Aria Mx system using cycling conditions mentioned in the protocol with annealing temperature of 58 degree Celsius.
Once master mix has been prepared and dispensed into individual wells and appropriate template (cDNA) has been added, allow the entire mix to stand for 5 minutes at room temperature for equalibration dye (fluorophore).
Multiplexed assay using IDT's prime time gene expression master mix performed very well. Repeated experiment three times with samples and running standards side-by-side. Got consistant Cq values in all three biological replicates with all three targets giving efficiency of 98% to 102 %.Quantification of gene expression of the target genes were closely correlated with those obtained in RNA-seq expreiments
Prepare your primers and probes at higher concentrations to store them as stock to prevent hydrolysis. Use IDT buffer recommended for resuspension of primers and probes.
Easy-to-use and less time on hands. Ideal for multiplexed assays
None
I recommend this product if one want to perform multiplexed qPCR assays. Technical support for trouble shooting from IDT is very excellentl