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