Direct Synthesis with Overlapping Oligos

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  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.