Oligo design tool for Guide-it SNP screening assays
Each SNP detection assay performed using the Guide-it Knockin Screening Kit involves the application of oligos designed to detect a specific substitution at a specified genomic locus. To simplify the oligo design process, we have developed the software tool below. Input the sequences corresponding to two alleles you wish to screen (e.g., wild-type/unedited and SNP/edited versions of your genomic target sequence), and the tool will output displacement, flap-probe, wild-type control, and SNP control oligos for your assay.
NOTE: This tool designs oligo probes for the screening of single-base substitutions only, not for insertions.
Design tool guidelines:
- Edited input sequences must include exactly one SNP.
- Both unedited and edited input sequences must be the same length.
- Input sequence lengths cannot exceed 1,000 bases.
- Input sequences should consist of at least 35 bases on either side of the SNP site. For optimal probe design, we recommend including ≥50 bases on either side.
NOTE: Please be careful to avoid the inclusion of extra spaces when inputting sequences, as these will be interpreted as part of the input and will likely return an error message.
To design oligos to detect multiple substitutions in parallel, please refer to "Detecting multiple edits" in the user guide.
Sequence before editing: GTCCGAGAACACCTTTATTGTGCACGTCCCCCGCAGAGCAGCCTCAGGCGTCCTGGTAGTAGTTGTTGAAGTTGATGCGCAGCAGAAAGT
Sequence after editing: GTCCGAGAACACCTTTATTGTGCACGTCCCCCGCAGAGCAGCCTCCGGCGTCCTGGTAGTAGTTGTTGAAGTTGATGCGCAGCAGAAAGT
Gene editing resources
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