Duke University/19 June 2009

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(New page: Did assembly PCR with different conditions (decreased extension time, increased number of cycles))
 
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Did assembly PCR with different conditions (decreased extension time, increased number of cycles)
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[[Team:Duke | Return ]]
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==June 19, 2009==
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*E: Assembly PCR (done by Maggie) with different conditions (decreased extension time, increased number of cycles)
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<pre>
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5x HF buffer 5 ul
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dNTPs 2 ul
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insert 7.18 ul
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Phusion DNA polymerase 0.4 ul
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H2O 10.42 ul
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25 ul
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98°C 30s 1x
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98°C 10s
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70-55°C 30s
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72°C 2m 40x
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72°C 5m
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4°C infinity 1x
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Fragment # Name Mass (ng) Volume (ul)
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1 pASKphaA 195.3 4.76
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2 phaB 41 0.60
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3 Termcat2 78.4 1.14
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4 phaC 86.4 0.68
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~400 7.14
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</pre>

Latest revision as of 16:46, 21 October 2009

Return

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
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