Davidson Missouri W/Primer dimer

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Generation of oligos and genes through PCR with Primer Dimers

1) Making sure that each primer does not contain substantial internal complementary sequences, divide the entire sequence into primers of no more than 70 nucleotides in length which have at least 12 bp overlap with the on either side.

2) Order these short oligos.

3) Run PCR with the first possible pair for each primer.

4) Gel purify to get the product you want for each segment.

5) Combine every other segment with the segment that would be immediately downstream.

6) repeat steps 3-5.