Difference between revisions of "Part:BBa K200001:Design"
(→‎Design Notes)
Line 15: Line 15:
 
<br>
 
<br>
 
<b>GGACTAGTA</b>TTATTTTTTCGCGGGTGAAAC<br>
 
<b>GGACTAGTA</b>TTATTTTTTCGCGGGTGAAAC<br>
 +
 +
 +
<i>BioBrick overhang shown in bold </i>
  
 
===Source===
 
===Source===

Revision as of 14:42, 29 September 2009

Dam methylase -> Dam


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 306
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The Dam methylase was obtained through PCR using BL21 as the genomic DNA template. Primers were designed to include overhangs coding for XbaI and SpeI recognition sites in order to allow the gene to be BioBricked according to the BioBrick Standard. The primers are as follows:

  • Forward PCR primer containing XbaI overhang (bold) for BioBricking:


GCTCTAGATGAAGAAAAATCGCGCTTTTTTG

  • Reverse PCR primer containing SpeI overhang (bold) for BioBricking:


GGACTAGTATTATTTTTTCGCGGGTGAAAC


BioBrick overhang shown in bold

Source

This is one of three site-specific DNA methylases found in most laboratory strains of E. coli.

References