Difference between revisions of "Part:BBa K777109:Design"

(→‎Design Notes)
(→‎Design Notes)
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* The ''fliC'' gene contained three PstI sites and one SpeI site that had to be removed. These mutations were induced with different primers via overlap-PCR.
 
* The ''fliC'' gene contained three PstI sites and one SpeI site that had to be removed. These mutations were induced with different primers via overlap-PCR.
 
* ''fliC'' was amplified from genomic DNA of ''E. coli'' DH10B via PCR using the following primers:  
 
* ''fliC'' was amplified from genomic DNA of ''E. coli'' DH10B via PCR using the following primers:  
−
** Fwd: gtttcttcgaattcgcggccgcttctagatggcacaagtcattaatacc
+
** Fwd: 5'-gtttcttcgaattcgcggccgcttctagatggcacaagtcattaatacc-3'
−
** Rev: gtttcttcgaattcgcggccgcttctagttaaccctgGagcagagacagaacc <br> (the capitalized "G" induces the mutation for removal of the third PstI site)
+
** Rev: 5'-gtttcttcgaattcgcggccgcttctagttaaccctgGagcagagacagaacc-3' <br> (the capitalized "G" induces the mutation for removal of the third PstI site)
  
 
===Source===
 
===Source===

Revision as of 08:45, 19 September 2012

fliC


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1224
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 301
    Illegal AgeI site found at 709
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

  • The fliC gene contained three PstI sites and one SpeI site that had to be removed. These mutations were induced with different primers via overlap-PCR.
  • fliC was amplified from genomic DNA of E. coli DH10B via PCR using the following primers:
    • Fwd: 5'-gtttcttcgaattcgcggccgcttctagatggcacaagtcattaatacc-3'
    • Rev: 5'-gtttcttcgaattcgcggccgcttctagttaaccctgGagcagagacagaacc-3'
      (the capitalized "G" induces the mutation for removal of the third PstI site)

Source

  • The part was amplified from genomic DNA of E. coli str. K-12 substr. DH10B, complete genome (CP000948.1).

References