Duke University/19 June 2009

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June 19, 2009

 * E: Assembly PCR (done by Maggie) with different conditions (decreased extension time, increased number of cycles)

5x HF buffer 5 ul dNTPs 2 ul insert 7.18 ul Phusion DNA polymerase 0.4 ul H2O 10.42 ul 25 ul

98°C 30s 1x 98°C 10s 70-55°C 30s 72°C 2m 40x 72°C 5m 4°C infinity 1x

Fragment # Name Mass (ng) Volume (ul) 1 pASKphaA 195.3 4.76 2 phaB 41 0.60 3 Termcat2 78.4 1.14 4 phaC 86.4 0.68 ~400 7.14