Duke University/18 June 2009

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Did assembly PCR <br>
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[[Team:Duke | Return]]
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Ran gels to check PCRs
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==June 18, 2009==
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*E: Assembly PCR, Gels to check PCRs  
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<pre>
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2x Phusion mix 12.5 ul
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insert 7.14 ul
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H2O 5.36 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 5s
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72°C 30s 25x
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72°C 2m
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4°C infinity 1x
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</pre>
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Fragment
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{| class="wikitable" border="1"
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! #
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! Name Size/Length (bp)
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! Concentration (ng/ul)
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! Mass (ng)
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! Volume (ul)
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|-
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| 1 || pASKphaA || 4215 || 41 || 194 || 4.73
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|-
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| 2 || phaB || 896 || 68 || 41 || 0.60
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|-
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| 3 || Termcat2 || 1701 || 69 || 78 || 1.13
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|-
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| 4 || phaC || 1872 || 126 || 86 || 0.68
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|-
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| ~400 || 7.14
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|}
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Length of plasmid: 8360 bp + 4 Histags * 6 aa * 3 bp = 8432 bp
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*R: Smear because of misformed gel, no band
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*C: Change conditions

Latest revision as of 16:45, 21 October 2009

Return

June 18, 2009

  • E: Assembly PCR, Gels to check PCRs 
2x Phusion mix 12.5 ul 
insert 7.14 ul 
H2O 5.36 ul 
25 ul 

98°C 30s 1x
98°C 10s
70-55°C 5s 
72°C 30s 25x
72°C 2m
4°C infinity 1x

Fragment

# Name Size/Length (bp) Concentration (ng/ul) Mass (ng) Volume (ul)
1 pASKphaA 4215 41 194 4.73
2 phaB 896 68 41 0.60
3 Termcat2 1701 69 78 1.13
4 phaC 1872 126 86 0.68
~400 7.14

Length of plasmid: 8360 bp + 4 Histags * 6 aa * 3 bp = 8432 bp

  • R: Smear because of misformed gel, no band
  • C: Change conditions
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