I start with IDT ordered 124 bp ssDNA that is PCR amplified, then I use a QIAgen agarose gel extraction to get my exact product, and I'm trying to purify ~95bp RNA by denaturing PAGE to prevent secondary structure formation, since I'm making an RNA hairpin. This process is very common in my lab: using "GGG" after promoter, promoter sequence checked, desalting the RNA product before T7 (I tried with and without this step-10kMWCO for 29kMW RNA), running O/N, followed by 1 hr DNase. Yet, still no bands at all for my sample but the control is still working (albeit very smeary) and I know it's not running off due to the ladder. Concentration verified by nanodrop.

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