Coding

Part:BBa_K4335009

Designed by: Hang Zhang   Group: iGEM22_UESTC-BioTech   (2022-10-10)
Revision as of 18:47, 10 October 2022 by Hang Zhang (Talk | contribs)


Insert site+gRNA scaffold

The transcript of gRNA scaffold can bind to SpCas9. The Insert site can be inserted into the sgRNA targeting specific target genes through the Golden Gate assembly.

Usage

Template of sgRNA. Use Golden Gate (BsaⅠ) to replace the template sequence with a ~20bp guide sequence. Add 'ACTT' on the 5'-terminal of your guide sequence and 'AAAC' on the reverse 5'-terminal.

sgRNA

Result

The following is the PCR verification electrophoresis diagram after we assembled various sgRNAs in Golden Gate Assembly.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 25
    Illegal BsaI.rc site found at 7


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Categories
Parameters
None