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  • What paramters should I use to create a strong positional (3') bias?
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  1. That depends on the protocol, a classical 5' > 3' bias is provoked by template loss during reverse transcription when carried out before fragmentation. Use the 5.3 Poly-dT Priming and DNAse Digestion (S.cerevisiae) template and consider to reduce the parameters RT_MIN and RT_MAX in order to make the bias effects stronger.