After delivery of the editing complex, you normally have a heterogeneous cell population consisting of unedited and edited cells. Edited cells may either be heterozygous (one allele edited) or homozygous (both alleles edited).
Depending on the goal of your experiment and your downstream analysis, you may either continue with a heterogeneous cell population or you may have to pick, analyse and expand individual edited cells.
You will find a number of assays and kits below which are useful for analysis of your edited cell population.
DNA isolation from edited cells
DNA isolation using QuickExtract
A simple, rapid extraction of PCR-ready DNA for screening your edited cells.
- 8-minute extraction protocol for most sample types
- No centrifugation steps or spin columns
- Automation-friendly: simple protocol integrates easily into automated workflows
- Safe: uses only non-toxic reagents
- Recommended for rapid, easy sample prep for CRISPR mutation detection assays
| Cat-No. | Item | Size | Price (CHF) |
|---|---|---|---|
| QE09050 | QuickExtract DNA Extraction Solution 1.0 | 50 ml | 525.00 |
DNA extraction using Norgen DNA extraction kits
DNA extraction kits from Norgen Biotek offer a rapid and convenient spin column procedure. Purified DNA is of the highest quality and integrity for sensitive downstream applications. The Cells and Tissue DNA Isolation Micro Kit is optimized for small inputs of cells and tissues, such as laser-captured microdissection (LCM).
Cas9 ELISA Assay
Cas9 ELISA Assay (Cas9-1000)
The assay can be used to monitor S. pyogenes Cas9 in cell culture lysate and is ideal for screening experimental samples utilizing the CRISPR/Cas9 gene editing system.
Microtitration wells coated with murine anti-Cas9 capture antibody are exposed to test specimens, which may contain Cas9 reactive determinants. The Cas9 antigen in the specimen is specifically captured onto the immobilized antibody during specimen incubation. The captured antigen is then reacted with a biotinylated mouse monoclonal Cas9 detection antibody. Subsequently, Streptavidin-HRP conjugate is then added. Following a wash cycle, specifically bound enzyme conjugate is detected by reaction with the Substrate Solution, tetramethylbenzidine (TMB). The assay is measured spectrophotometrically to indicate the level of Cas9 reactive determinants present in a sample. The assay can be used to monitor S. pyogenes Cas9 in cell culture lysate and is ideal for screening experimental samples utilizing the CRISPR/Cas9 gene editing system
Alt-R® Genome Editing Detection Kit
For detection of on-target or known off-target CRISPR events in cultured cells. This T7 endonuclease I (T7EI) mismatch cleavage assay detects on-target genome editing and estimates genome editing efficiency in CRISPR- treated cells.
- Fast — does not require purification of PCR products before analysis
- Consistent — analyse easy-to-read electrophoresis results
- Quantifiable — gel band intensities relate to genome editing efficiency
- Versatile — convenient for a single sample or high-throughput analysis in plate format
| Cat-No. | Kit | Size |
|---|---|---|
| 1075931 | Alt-R® Genome Editing Detection Kit | 25 rxn |
rhAmpSeq for evaluation of edits
The rhAmpSeq system enables highly accurate amplicon sequencing on GenapSysTM and IlluminaTM next- generation sequencing (NGS) platforms. The fast and easy rhAmpSeq workflow, based on IDT’s proprietary RNase H2-dependent PCR technology, generates NGS-ready amplicon libraries for deep, targeted resequencing.
- Achieve cost-effective library preparation using custom or predesigned panels
- Reduce the formation of primer dimers or misprimed PCR products
- Perform only two PCR amplification steps in a fast, easy workflow
| Supplier | Item | Size |
|---|---|---|
| IDT | Custom rhAmpSeq Panels for CRISP analysis | 1 panel |
Storage of edited cell pools and clonal lines
Store your valuable cell pools and clonal cell lines using BambankerTM in order to ensure maximum viability during long-term storage. Its innovative formulation simplifies the freezing process and greatly increases cell viability. With BambankerTM, the viability of cells during freeze/thaw cycles is greatly enhanced, even for fragile cells such as embryonic stem cells.
For research use only. Not for use in diagnostic procedures. Unless otherwise agreed to in writing, IDT does not intend for these products to be used in clinical applications and does not warrant their fitness or suitability for any clinical diagnostic use. Purchaser is solely responsible for all decisions regarding the use of these products and any associated regulatory or legal obligations.