Difference between revisions of "Part:BBa K200004:Design"

(→‎Design Notes)
(→‎Design Notes)
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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.
 
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:
 
The primers are as follows:
 +
*Forward PCR primer containing XbaI overhang (bold) for BioBricking:<br>
 +
<br>
 +
<b>GCTCTAG</b>ATGAAGAAAAATCGCGCTTTTTTG<br>
 +
<br>
 +
*Reverse PCR primer containing SpeI overhang (bold) for BioBricking:<br>
 +
<br>
 +
<b>GGACTAGTA</b>TTATTTTTTCGCGGGTGAAAC<br>
  
 
===Source===
 
===Source===

Revision as of 11:18, 28 August 2009

TaqI


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 18
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 157


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