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Abstract: This protocol explains how to prepare dsDNA and ssRNA target positive control for LAMP and RT-LAMP reactions. Particularly it describes the pipeline for SARS-Cov-2 Nucleocapsid (N) gene positive control which is included in the ReClone collection. The general procedure involves amplifying the gene of interest with primers which add T7 promoter and T7 terminator to its sequence followed by T7 in vitro transcription to obtain the ssRNA. Sequence elaboration: Sequence OrfN_5 was designed and synthesized based on IDT SARS-CoV-2 positive control plasmid (CAT#10006625_2019).