Part:BBa_K4409015
gRNA (AAG+ BBa_K3859000)
This is a gRNA used in a CRISPR-Cas12 system. It mainly comprises a Cas12 handle (BBa_K2961003) which combines with a Cas12 enzyme and a guide sequence (BBa_K3859000) which binds with a dsDNA to activate the Cas12 enzyme.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage and Results
We design a plasmid based on pET-28a plasmid by t-PCR method. We inserted the targeted sequence between the restriction enzyme site TCTAGA and CTCGAG. The PCR results show the successful plasmids construction since the site of the band is between 5000bp and 8000bp, shown in Graph 1. And the length of plasmid we design is also inside this region, exactly 5420bp. Moreover, we confirmed the sequence by gene sequencing, shown in Graph 2. All of the base pairs in the main region turns out to be accurate.
The igRNA produced can also produce fluorescence which is significantly bigger than negative control groups. The fluorescence produced is listed in the following graph:
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