Product Details
(This product is Patent Pending)
Get faster production of CRISPR edited strains - use our Return-to-N2 Co-CRISPR Screening Kit for your next C. elegans transgenic.
Co-CRISPR allows you to quickly identify C. elegans with a high likelihood of having your favorite genomic edit, but current methods (markers with dpy-10, sqt-1, etc) require additional screening to remove the marker. NemaMetrix’s Return-to-N2 Co-CRISPR Screening Kit allows you to use restoration of wild type to speed your discovery of the target site edit. A simple PCR screen in your crawling F1 founders finds your target site heterozygote. Then the same PCR screen of F2 progeny finds the homozygous line. The unc-18 PCR test that is included in the kit can also be used to confirm that the unc-18 locus is wild-type, or you can check the F3 progeny for the absence of the unc-18 phenotype to confirm that your line is free of marker background.
Advantages
- Restore unc-18 to wild type as replacement for the dpy-10 screen.
- Crawling lines routinely have over 50% likelihood of having your target gene edit.
- No marker phenotype to remove after desired edit is found.
- Saves anywhere from ~1.5-10 hours of hands-on time and ~2-20 days of time overall.
Use this strain with any genomic edit without taking into consideration the distance between the co-CRISPR locus and the locus you are editing. Using a return-to-N2 co-CRISPR eliminates the chance your edit will be linked to the co-CRISPR mutant phenotype.
The Co-CRISPR Screening Kit with unc-18 includes reagents for 3 experiments:
- unc-18 co-CRISPR strain
- CRISPR injection mix base (includes unc-18 sgRNA and repair template)
- PCR primer mix to verify return-to-N2 sequence
*Not included: Cas9, sgRNAs and donor-homology for your target edit
Need help designing an edit?
No problem. Feel free to reach out to us at support@nemametrix.com
Reagents that work well with this kit?
Storage
- This kit is shipped at room temperature.
- For COMPONENT A (Injection Mix) at -20C upon receipt
- For COMPONENT D (NGM plate with worms) store between 15 and 20C
- For COMPONENT E (Screening Primers) store at -4C
Documents
Protocol (PDF)
Safety Data Sheet (PDF)
Quick Reference Card
Frequently Asked Questions
- This kit is shipped at room temperature.
- For COMPONENT A (Injection Mix) at -20C upon receipt
- For COMPONENT D (NGM plate with worms) store between 15 and 20C
- For COMPONENT E (Screening Primers) store at -4C
The kit shelf life has been tested up to 4 weeks, we will continue to update the shelf life as testing progresses.
The unc-18 strain provided in this kit has an early stop that disrupts the protein's function. Our co-CRISPR kit returns this locus back to wildtype, restoring the gene's function.
The kit contains what you need to make the co-CRISPR edit. You will still need the Cas9, sgRNA(s), and ODN to make your desired edit.
We recommend what we use in our own labs and that is our PCR Master Mix and Lysis Kit
We recommend PNA Bio Cas9 CP02
It is not. This co-CRISPR is designed for C. elegans only.
It is not. This co-CRISPR kit is designed for C. elegans only.
The co-CRISPR locus is on chromosome X.
Animals that are heterozygous for the co-CRISPR will crawl like wild type animals. However, unlike animals that are homozygous for the co-CRISPR edit, they will have both crawling and unc progeny.
No, we have not seen off-target editing with these co-CRISPR components.
The NemaMetrix scientists are experts at designing edits and we are more that happy to help. For additional information or questions, please contact support@nemametrix.com
Yes.
The NemaMetrix Team is always here to help you. For additional information or questions, please contact support@nemametrix.com
We always want to make our customers happy and if there is something that you want, we are here for you! Please contact our team to discuss your needs at support@nemametrix.com
The allele name is knu409
The strain number is COP1403