Universal Probe Library Assay
Based on the Universal Probe Library Assay Quick Guide from Roche
Design your primers
- Each gene you analyze requires a forward primer, reverse primer, and a UPL probe.
- Use Roche's Assay Design Center to design optimal primers and identify the right probe for your gene(s) of interest.
- The forward and reverse primers need to be ordered from a DNA synthesis company (e.g., IDT DNA, Promega, etc.), and the UPL oligo comes from Roche.
Design your experiment
How many reactions should I plan to run? Each experimental sample is a template. The gene being detected is often referred to as a target. Each unique template and target combination requires its own reaction. You will also need to set up a no template control to observe the amount of background noise from that reaction. For instance, a scientist wants to measure differences the expression of genes A, B, and C in an experiment where cells were treated with a drug, or untreated. All of the unique reactions she must set up are:
|Rxn 1:||treated cells||gene A, primer set A|
|Rxn 2:||treated cells||gene B, primer set B|
|Rxn 3:||treated cells||gene C, primer set C|
|Rxn 4:||untreated cells||gene A, primer set A|
|Rxn 5:||untreated cells||gene B, primer set B|
|Rxn 6:||untreated cells||gene C, primer set C|
|Rxn 7:||no template||gene A, primer set A|
|Rxn 8:||no template||gene B, primer set B|
|Rxn 9:||no template|| gene C, primer set C