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• 18:04, 29 May 2025 ESO wikiadmin talk contribs created page Design principles for PCR primers used for amplification and cloning of cDNA (Created page with "'''Restriction Enzyme Sites:''' For conventional cloning with restriction enzymes, the recognition sites are added to the 5’ end of the primer. Check that the involved '''restriction enzymes do NOT cut within the cloned cDNA''' and that they do '''NOT cut the vector outside the intended position'''. Make sure to preserve the reading frame, if the DNA is cloned 3’ or 5’ of existing ORFs (e.g. GFP) in the vector. Add 3 to 4 meaningless nucleotides (e.g. ATAA-) at the...") Tag: Visual edit