Difference between revisions of "Part:BBa K4615000:Design"

 
Line 9: Line 9:
 
We obtained this part by introducing 20 bp of sgRNA within the tpiA genome that guides the Cas9 protein to cut and create strand breaks.
 
We obtained this part by introducing 20 bp of sgRNA within the tpiA genome that guides the Cas9 protein to cut and create strand breaks.
  
−
 
+
Melting temp: 62C
 +
GC 42%
  
 
===Source===
 
===Source===

Revision as of 15:56, 12 October 2023


Forward primer to insert 20 bp of sgRNA of tpiA into pTargetF


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 27
    Illegal XbaI site found at 33
    Illegal PstI site found at 45
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 27
    Illegal PstI site found at 45
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 27
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 27
    Illegal XbaI site found at 33
    Illegal PstI site found at 45
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 27
    Illegal XbaI site found at 33
    Illegal PstI site found at 45
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We obtained this part by introducing 20 bp of sgRNA within the tpiA genome that guides the Cas9 protein to cut and create strand breaks.

Melting temp: 62C GC 42%

Source

The 20bp of sgRNA was generated using CHOPCHOP and analyzed based on various factors such as GC content, self-complementarity and efficiency.

References