Team:Stockholm/20 September 2010

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(New page: {{Stockholm/Top2}} ==Andreas==)
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{{Stockholm/Top2}}
{{Stockholm/Top2}}
==Andreas==
==Andreas==
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===Assembly of new parts===
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 +
#'''pSB1K3.N-LMWP⋅SOD⋅His'''
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#*Dig pSB1C3.N-LMWP (E+A)
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#*Dig pMA.SOD⋅His (N+P)
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#*Dig pSB1K3.RFP (E+P)
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#'''pSB1C3.N-LMWP⋅SOD⋅His'''
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#*Dig pSB1C3.N-LMWP (A+S)
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#*Dig pMA.SOD⋅His (N+S)
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 +
====Digestions====
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{|border="1" cellpadding="1" cellspacing="0"
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| 
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!width="50"|pSB1C3. N-LMWP
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!width="50"|pMA. SOD⋅His
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!width="50"|pSB1C3. N-LMWP
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!width="50"|pSB1K3. N-TAT⋅SOD⋅ His 4
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|-
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|10X FastDigest buffer
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|align="center"|3
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|align="center"|3
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|align="center"|3
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|align="center"|2
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|-
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|dH<sub>2</sub>O
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|align="center"|15.2
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|align="center"|4.1
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|align="center"|15.2
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|align="center"|11.4
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|-
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|DNA (1 &mu;g)
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|align="center"|9.8
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|align="center"|20.9
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|align="center"|9.8
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|align="center"|4.6
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|-
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|AgeI
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|align="center"|1
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|align="center"|0
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|align="center"|1
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|align="center"|0
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|-
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|NgoMIV
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|align="center"|0
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|align="center"|1
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|align="center"|0
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|align="center"|0
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|-
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|FD SpeI
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|align="center"|1
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|align="center"|1
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|align="center"|0
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|align="center"|0
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|-
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|FD EcoRI
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|align="center"|0
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|align="center"|0
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|align="center"|1
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|align="center"|0
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|-
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|FD PstI
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|align="center"|0
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|align="center"|0
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|align="center"|0
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|align="center"|1
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|-
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|FD XbaI
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|align="center"|0
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|align="center"|0
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|align="center"|0
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|align="center"|1
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|-
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|&nbsp;
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!30 &mu;l
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!30 &mu;l
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!30 &mu;l
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!20 &mu;l
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|}
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*Incubation: 37 &deg;C, 2:00 (NgoMIV & AgeI); 0:30 (FD)
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*Inactivation: 80 &deg;C, 20 min

Revision as of 22:06, 21 September 2010

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Andreas

Assembly of new parts

  1. pSB1K3.N-LMWP⋅SOD⋅His
    • Dig pSB1C3.N-LMWP (E+A)
    • Dig pMA.SOD⋅His (N+P)
    • Dig pSB1K3.RFP (E+P)
  2. pSB1C3.N-LMWP⋅SOD⋅His
    • Dig pSB1C3.N-LMWP (A+S)
    • Dig pMA.SOD⋅His (N+S)

Digestions

  pSB1C3. N-LMWP pMA. SOD⋅His pSB1C3. N-LMWP pSB1K3. N-TAT⋅SOD⋅ His 4
10X FastDigest buffer 3 3 3 2
dH2O 15.2 4.1 15.2 11.4
DNA (1 μg) 9.8 20.9 9.8 4.6
AgeI 1 0 1 0
NgoMIV 0 1 0 0
FD SpeI 1 1 0 0
FD EcoRI 0 0 1 0
FD PstI 0 0 0 1
FD XbaI 0 0 0 1
  30 μl 30 μl 30 μl 20 μl
  • Incubation: 37 °C, 2:00 (NgoMIV & AgeI); 0:30 (FD)
  • Inactivation: 80 °C, 20 min
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