Difference between revisions of "Part:BBa K1539002:Design"

(→‎Design Notes)
(→‎Design Notes)
 
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===Design Notes===
 
===Design Notes===
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mCherry from BBa_J0650 was amplified with BBa_K1539034.  This product were Dpn1 digested and then PCR cleaned up.  The product was then PCR re-amplified with BBa_K1539089, digested with XbaI and PstI and ligated into a similarly digested pSB1c3 backbone.  Amplification of each step was confirmed by gel electrophresis producing bands approximately 900-950 bp (~700 bp for mCherry, 30 bp for RBS, 60 bp for promoter, and ~150 in extra sequence added by codons beyond the suffix to the reverse sequencing primer).
+
mCherry from BBa_J0650 was amplified with BBa_K1539034.  This product was Dpn1 digested and then PCR cleaned up.  The product was then PCR re-amplified with BBa_K1539089, digested with XbaI and PstI and ligated into a similarly digested pSB1c3 backbone.  Amplification of each step was confirmed by gel electrophresis producing bands approximately 900-950 bp (~700 bp for mCherry, 30 bp for RBS, 60 bp for promoter, and ~150 in extra sequence added by codons beyond the suffix to the reverse sequencing primer, VR).
  
 
===Source===
 
===Source===

Latest revision as of 20:59, 1 November 2014


LPromoter->LRBS->mCherry


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

mCherry from BBa_J0650 was amplified with BBa_K1539034. This product was Dpn1 digested and then PCR cleaned up. The product was then PCR re-amplified with BBa_K1539089, digested with XbaI and PstI and ligated into a similarly digested pSB1c3 backbone. Amplification of each step was confirmed by gel electrophresis producing bands approximately 900-950 bp (~700 bp for mCherry, 30 bp for RBS, 60 bp for promoter, and ~150 in extra sequence added by codons beyond the suffix to the reverse sequencing primer, VR).

Source

mCherry from BBa_J06504 RBS and promoter from created primers

References