I am trying to do a Sanger sequencing on my miRNA, and look into specific seed region of editing. However, my primers are derived from primary miRNA, and I do know pri-miRNAs are relatively low in abundance, thus it prompts me to do a pre-amp steps in order for the sequencing to go correctly. Does anyone have any idea/ or inpur in regards to this matter? Some people suggested nested PCR but I have little knowledge about that technique.

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