Ultramers® and miniGenes™ as positive controls
Proper controls are a crucial part to any qPCR experiment. IDT recommends that you include both positive and negative controls when designing and running qPCR reactions. Examples of negative controls include no-template, no-reverse transcriptase, and no-amplification controls. Examples of positive controls include exogenous and endogenous target controls.
An exogenous positive control is external DNA or RNA carrying a target of interest. This type of control reaction will alert you to any components in the sample that might inhibit reverse transcription and/or PCR. IDT synthesizes miniGenes™ and Ultramer® Oligonucleotides which can serve the role of an exogenous positive control. Ultramer Oligonucleotides are high-fidelity long oligos (up to 200 bases) and miniGenes are double-stranded DNAs of up to 400 bp that are constructed using Ultramers. These nucleic acids provide a known starting copy number and so serve as standards with known concentrations. In addition, we recommend adding a phage promoter region (such as T7 or SP6) to the amplicon sequence. This will generate an in vitro transcribed RNA to provide a control for the reverse transcription step.
Ultramer Oligonucleotides are also suitable for demanding applications like cloning and gene construction, and can save researchers a great deal of time and trouble through direct synthesis of the entire fragment. miniGenes are part of a confidential and guaranteed gene synthesis service. All miniGenes are sequence verified, and delivered in a purified plasmid.