Difference between revisions of "Part:BBa K4615000:Design"
 
Line 10: Line 10:
  
 
Melting temp: 62C
 
Melting temp: 62C
 +
 
GC 42%
 
GC 42%
  

Latest 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