Is DNase treatment recommended for single cell RT-PCR?
As the AmpliGrid slide shows an outstanding sensitivity it is generally possible to amplify single DNA copies during an RT-PCR experiment. During the analysis of high expressed genes the final amount from the DNA amplification can be neglected compared to the RNA amplification whereas analyzing medium or low expressors the amplified DNA might interfere with the results. We recommend two strategies to overcome that issue: (1) Primerdesign: Use intron spanning primers to be able to distinguish between DNA and RNA amplicons. In case you need assistance for designing the appropriate primers please make use of our service. (2) DNase treatment: if an intron spanning primerdesign is not possible please integrate a DNase step into your RT setup (see here for protocol). Please note: any additional step during a single cell experiment is prone to sample loss or degradation of RNA.
