Team:University College London/Protocols/PrimerDesign

Primer Design

Primers should be designed to: 1) Be 18-25 nucleotides in length 2) Have a melting temperature that does not differ by more than 5oC between the two primers of a pair 3) GC nucleotides should constitute 40-60% 4) Have a 3’primer end that is not complimentary to any region of the primer pair 5) Contain an equal distribution of the nucleotides 6) Closely match the target sequence (and so known polymorphic sites are avoided) 7) Have a GC clamp on either end