I have know sequence of mRNA. but i need to amplify the unknown region of 5'end ? to do this that needs GSP1(reverse) and GSP2 primer(forward). my kit protocol states your GSP2 primer is anneals to sequence located 3' (with respect to cDNA not mRNA). here how I design the GSP2 ? is from directedly the mRNA or from its complement ? for more clarity I attached my kit protocol and if there is any volunteer explain page. 4 from the document ? thanks in advance