Difference between revisions of "Part:BBa K4223003"
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<partinfo>BBa_K4223003 short</partinfo> | <partinfo>BBa_K4223003 short</partinfo> | ||
| − | Cas13a tolerates one or two mismatches between crRNA and the target sequence, which will result in a greatly reduced cleavage efficiency of Cas13a protein, and this wrong cleavage will also lead to "false positives" in the detection. | + | Cas13a tolerates one or two mismatches between crRNA and the target sequence, which will result in a greatly reduced cleavage efficiency of Cas13a protein, and this wrong cleavage will also lead to "false positives" in the detection. <br> |
| − | In order to confirm the occurrence of false positives, Hainanu-China designed a series of sequences with single nucleotide polymorphisms (SNPS) based on the target RNA, which were detected by fluorescence reporter. | + | In order to confirm the occurrence of false positives, Hainanu-China designed a series of sequences with single nucleotide polymorphisms (SNPS) based on the target RNA, which were detected by fluorescence reporter. <br> |
It should be noted that fluorescence was generated by trans-cleavage of ssRNA cleavage Reporter after Cas13a recognized and cleaved the target RNA. | It should be noted that fluorescence was generated by trans-cleavage of ssRNA cleavage Reporter after Cas13a recognized and cleaved the target RNA. | ||
Revision as of 08:39, 20 September 2022
Transcription system of target RNA for LbCas13a
Cas13a tolerates one or two mismatches between crRNA and the target sequence, which will result in a greatly reduced cleavage efficiency of Cas13a protein, and this wrong cleavage will also lead to "false positives" in the detection.
In order to confirm the occurrence of false positives, Hainanu-China designed a series of sequences with single nucleotide polymorphisms (SNPS) based on the target RNA, which were detected by fluorescence reporter.
It should be noted that fluorescence was generated by trans-cleavage of ssRNA cleavage Reporter after Cas13a recognized and cleaved the target RNA.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Unknown
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 100
- 1000COMPATIBLE WITH RFC[1000]