I am currently working on LncRNA; to know the lncRNA-protein interactions I want to do RNA pull down assay, so I need to design primers with T7 promoter. I need assistance in this regard.
Nothing very complicated, just add the sequence of the T7 promoter upstream of the specific sequence of your forward primer. Adding a few nucleotides before the promoter can help with the transcription since it will help with the polymerase loading, I usually add 4-5 As. For the pulldown we preform the IVT with biotinylated UTP using a mix of regular and labeled UTP at a ratio that will give us 5-6 biotinylated Us per transcript.