Difference between revisions of "Part:BBa I751300:Design"

 
(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
 +
1. Cut Biobrick part by EcoRI and SpeI.
 +
2. Cut pBR322 by EcoRI and NheI.
 +
3. Ligation and TF.
  
  
 +
pBR322 has NheI site downstream of EcoRI site.
  
 +
NheI sequence is "G^CTAGC", and SpeI sequence is "A^CTAGT". They have the sequence "CTAG" in common. So, NheI fragment and SpeI fragment can be ligated. However, this procedure kills SpeI site, and you can't assemble parts anymore. (EcoRI and XbaI site is still alive, so you can assemble upward)
  
 +
 +
This procedure allowed us to introduce any Biobrick part into pBR322 plasmid.
  
 
===Source===
 
===Source===

Revision as of 15:18, 25 October 2007


pBR322 luxI


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 655
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

1. Cut Biobrick part by EcoRI and SpeI. 2. Cut pBR322 by EcoRI and NheI. 3. Ligation and TF.


pBR322 has NheI site downstream of EcoRI site.

NheI sequence is "G^CTAGC", and SpeI sequence is "A^CTAGT". They have the sequence "CTAG" in common. So, NheI fragment and SpeI fragment can be ligated. However, this procedure kills SpeI site, and you can't assemble parts anymore. (EcoRI and XbaI site is still alive, so you can assemble upward) 


This procedure allowed us to introduce any Biobrick part into pBR322 plasmid.

Source

References