Difference between revisions of "Part:BBa K200001:Design"

 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K200001 short</partinfo>
 
<partinfo>BBa_K200001 short</partinfo>
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===Design Notes===
 
===Design Notes===
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None
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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.
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The primers are as follows:
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*Forward PCR primer containing XbaI overhang (bold) for BioBricking:<br>
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<br>
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<b>GCTCTAG</b>ATGAAGAAAAATCGCGCTTTTTTG<br>
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<br>
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*Reverse PCR primer containing SpeI overhang (bold) for BioBricking:<br>
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<br>
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<b>GGACTAGTA</b>TTATTTTTTCGCGGGTGAAAC<br>
  
 
===Source===
 
===Source===

Revision as of 14:36, 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

Source

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

References