Gator Bio's GeneSwift Assay Kit is a simple, fast, and reliable method to determine the titer. It streamlines a two-step procedure involving both AAV lysis and genome hybridization within a single tube, succeeded by biolayer interferometry (BLI) detection.
Probe Design Parameters
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Basic Oligo Probe Design Instructions
Pick target sequence(s) anywhere (except ITR) in the insert or for genome integrity from the ends of the insert, e.g. CMV enhancer on one end and SV-40 at the other.
- Design one fluorescein-labeled probe and one biotin-labeled probe that hybridizes to the same strand of the target sequence (positive or negative).
- Both probes are approximately 35–40 nt long and should not overlap.
- The fluorescein-labeled probe is labeled only on the 5′ end.
- The biotin-labeled probe has an additional 30 Thymines (T) along with biotins added at both 5′ and 3′ ends.
- The calculated energy for the probes should be higher than −10 kcal/mol to minimize homo and heterodimer association.
- Linguistic sequence complexity (LC%) measures the vocabulary richness of a genetic text; 100% = maximum diversity.
Input & Results
- A name is not required but preferred for each probe (e.g. Seq1).
- Paste your target sequence in FASTA format. Only standard IUPAC nucleic acid characters are accepted.
- Use
[and]to specify each sequence location for probe design; use/…/to exclude regions.
Both forward (FWD) and reverse (RVS) oligos for the negative and positive strands are reported in paired format. Design a few different pairs to find the most optimal for your assay.