Difference between revisions of "Part:BBa K200017:Design"

 
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K200017 short</partinfo>
 
<partinfo>BBa_K200017 short</partinfo>
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===Design Notes===
 
===Design Notes===
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N/A
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OtsB ([https://parts.igem.org/Part:BBa_K200006 BBa_K200006]) was obtained from PCR from Escherichia Coli using BL21(DE3) as the genomic DNA template and the Pfu Ultra II enzyme. 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:
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<br>
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<br>
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Forward PCR primer containing XbaI overhang (bold) for BioBricking: <br>
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<b>GCTCTAG</b>ATGACAGAACCGTTAACCG <br>
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Recomended Temperatures for PCR : 54.5C(without overhang) and 64.8C(with overhang)
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<br><br>
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Reverse PCR primer containing SpeI overhang (bold) for BioBricking: <br>
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<b>GGACTAGTA</b>TTAGATACTACGACTAAACGAC<br>
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Recomended Temperatures for PCR : 54.7C(without overhang) and 64.2C(with overhang)<br><br>
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<i>Biobrick Overhang shown in Bold</i>
  
  
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===Source===
 
===Source===
  
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PCR of OtsB from Escherichia Coli BL21(DE3, ligated to RBS extracted from Registry
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PCR of OtsB from Escherichia Coli BL21(DE3), ligated to RBS extracted from Registry
  
 
===References===
 
===References===

Latest revision as of 21:47, 19 October 2009

RBS + OtsB


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 473
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

OtsB (BBa_K200006) was obtained from PCR from Escherichia Coli using BL21(DE3) as the genomic DNA template and the Pfu Ultra II enzyme. 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:
GCTCTAGATGACAGAACCGTTAACCG
Recomended Temperatures for PCR : 54.5C(without overhang) and 64.8C(with overhang)

Reverse PCR primer containing SpeI overhang (bold) for BioBricking:
GGACTAGTATTAGATACTACGACTAAACGAC
Recomended Temperatures for PCR : 54.7C(without overhang) and 64.2C(with overhang)

Biobrick Overhang shown in Bold


Source

PCR of OtsB from Escherichia Coli BL21(DE3), ligated to RBS extracted from Registry

References