Difference between revisions of "Part:BBa K1076001:Design"

(→‎Design Notes)
(→‎Design Notes)
Line 6: Line 6:
  
 
===Design Notes===
 
===Design Notes===
−
To upstream gene we used primers with Apa1(5') and NdeI(3') and to downsteam gene we used primers with NdeI(5') and BamH1(3')ends
+
 
 +
To design the upstream gene we used primers with Apa1(5') and NdeI(3') and to downsteam gene we used primers with NdeI(5') and BamH1(3')ends, this parts have Nde1 complementary site and they were ligate together. This method garantee generation of a deleted copy of the target gene using a two step assymmetric/crossover PCR amplification. The target in this case is the FadR.
  
 
===Source===
 
===Source===

Revision as of 23:28, 27 September 2013

Flanking genes for FadR deletion


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 768
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

To design the upstream gene we used primers with Apa1(5') and NdeI(3') and to downsteam gene we used primers with NdeI(5') and BamH1(3')ends, this parts have Nde1 complementary site and they were ligate together. This method garantee generation of a deleted copy of the target gene using a two step assymmetric/crossover PCR amplification. The target in this case is the FadR.

Source

It comes from amplified genomic sequence

References