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.Result
The following is the PCR verification electrophoresis diagram after we assembled various sgRNAs in Golden Gate Assembly.Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 25
Illegal BsaI.rc site found at 7
[edit]
Categories
Parameters
None |