Direct Synthesis with Overlapping Oligos
From GcatWiki
Design oligos with biobrick suffix, prefix, and at least 15 bp overlap.
Assemble reaction for PCR.
1 ul (10 ug) of oligo A
1 ul (10)ug) of oligo B
4 ul 10X PCR buffer w/ 15 mM MgCl2
2 ul 10X dNTDs
1 ul Taq
31 ul dH2O
5 cycles: 94 C 15 sec.; annealing temp. 15 sec.; 74 C 30 sec.
annealing temp. must be 4 C lower than Tm of either oligo.