I want to use one step RT-PCR to amplify human RB1 gene (mRNA-->cDNA). When I looked at NCBI nucleotide database (NM_000321.2), the gene has CDS on it. Which one should I amplify, the complete mRNA (4.7 kb) or CDS only (2.7 kb) ?
NB: I want to clone the gene to plasmid vector (vector size: 4.7 kb) in E. coli, then express the gene into protein