Difference between revisions of "Part:BBa K1431824:Design"
(→‎Design Notes)
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This Biobrick is constructed through 3 steps:
 
This Biobrick is constructed through 3 steps:
  
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# Do double enzyme digestion (XbaI & PstI for BBa_K1033916, SpeI & PstI for BBa_B0034) and do gel extraction, ligation, transformation, picking up single colonies, LB broth incubation, plasmid extraction and gel electrophoresis verification.
+
# Do double enzyme digestion (XbaI & PstI for BBa_K1033916, SpeI & PstI for BBa_B0031) and do gel extraction, ligation, transformation, picking up single colonies, LB broth incubation, plasmid extraction and gel electrophoresis verification.
−
# Do double enzyme digestion (XbaI & PstI for Step 1 product, SpeI & PstI for J23100) and do gel extraction, ligation, transformation, picking up single colonies ('''can select the positive colonies from the colony color'''), LB broth incubation, plasmid extraction and gel electrophoresis verification.
+
# Do double enzyme digestion (XbaI & PstI for Step 1 product, SpeI & PstI for J23106) and do gel extraction, ligation, transformation, picking up single colonies ('''can select the positive colonies from the colony color'''), LB broth incubation, plasmid extraction and gel electrophoresis verification.
 
# Do double enzyme digestion (EcoRI-HF & SpeI for Step 2 product, EcoRI-HF & XbaI for B0015) and do gel extraction, ligation, transformation, picking up single colonies ('''can select the positive colonies from the colony color'''), LB broth incubation, plasmid extraction and gel electrophoresis verification.
 
# Do double enzyme digestion (EcoRI-HF & SpeI for Step 2 product, EcoRI-HF & XbaI for B0015) and do gel extraction, ligation, transformation, picking up single colonies ('''can select the positive colonies from the colony color'''), LB broth incubation, plasmid extraction and gel electrophoresis verification.
 
# Cryopreserved the rest bacteria broth and send samples for sequencing.
 
# Cryopreserved the rest bacteria broth and send samples for sequencing.
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−
 
  
 
===Source===
 
===Source===

Revision as of 03:01, 18 October 2014

amajLime, yellow-green chromoprotein reporter system (Weak Promoter, Weak RBS)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This Biobrick is constructed through 3 steps:

  1. Do double enzyme digestion (XbaI & PstI for BBa_K1033916, SpeI & PstI for BBa_B0031) and do gel extraction, ligation, transformation, picking up single colonies, LB broth incubation, plasmid extraction and gel electrophoresis verification.
  2. Do double enzyme digestion (XbaI & PstI for Step 1 product, SpeI & PstI for J23106) and do gel extraction, ligation, transformation, picking up single colonies (can select the positive colonies from the colony color), LB broth incubation, plasmid extraction and gel electrophoresis verification.
  3. Do double enzyme digestion (EcoRI-HF & SpeI for Step 2 product, EcoRI-HF & XbaI for B0015) and do gel extraction, ligation, transformation, picking up single colonies (can select the positive colonies from the colony color), LB broth incubation, plasmid extraction and gel electrophoresis verification.
  4. Cryopreserved the rest bacteria broth and send samples for sequencing.

Source

will be added later

References