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