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Give us a vial of DNA and we will give you a vial of flies with the DNA integrated!
Drosophila transgenesis is a labour intensive process and requires optimised protocols and technical expertise. This process often diverts attention from the core research project. We are offering Drosophila Transgenesis services at competitive prices. We are offering both P-element mediated and PhiC31-integrase mediated transgenesis services. With the state-of-art equipment and the experienced staff we are able to generate transgenics in throughput rate for large scale projects.
1. PhiC31integrase mediated transgenesis:
Site-Specific Excel - Injection of user generated DNA into upto 300 embryos of flies containing PhiC31 integrase & selected attP docking site (click here and choose attP sites), crossing of G0 flies, screening the transformants and balancing the insertions.
Site-Specific BAC- This service includes injecting the BACs or fosmids (>30kb) into 500 embryos of strain containing PhiC31-integrase and an AttP docking site of the user’s choice and scoring the transformants, balancing and shipping.
Site-Specific CRISPR- This service includes injecting the plasmids with CRISPR/Cas9 guide RNA plasmid into upto 300 embryos of strain containing PhiC31-integrase and a docking site of the user’s choice and scoring the transformants, balancing and shipping.
2. CRISPR/Cas9 mediated mutagenesis:
gRNA design and synthesis- This service includes design of a single gRNA and its synthesis in a plasmid vector available in the facility (pBFVU6.2 or pCFD). User will get the plasmid DNA and the sequencing data.
Testing gRNA pairs in embryos- This service include injection of gRNA pairs in 50 embryos of Act5c-Cas9 transgenic, and PCR based screening for the efficiency of the pair.
Large (2kb+ deletions)- This service includes injecting dual-gRNA in ~300 embryos from appropriate Cas9 flies, G0 crosses, PCR based screening, sequence confirmation and balancing mutant flies.
Homologous recombination construct design and synthesis- This service includes design and synthesis of HR construct by Gibson Assembly. User will get the final construct and the sequencing data.
CRISPR based homologous recombination- This service includes injection of gRNA and HR construct provided by the user, crossing G0 flies and screening for visible markers and balancing the positive progeny.
3. Embryo Microinjection service:
This rapid mode service includes injection of user supplied DNA in 300 embyos in Standard Drosophila stocks.
*Injection in Non-standard stocks for any of the services may be requested. This additional service will require stock expansion and will be charged extra.
4. P-element mediated transgenesis:
P-element classic- Injection of DNA into w1118 embryos, raising the survived larvae to adulthood and crossing the G0 flies (>100 individual crosses). Identification and collection of transformants; the collected transformant flies (at least 3 independent lines) are sent to the customer and original vials are maintained in the facility for two weeks as backup.
P-element excel- The service includes mapping of the transformants obtained in addition to service in P-element classic.
5. Custom services are also available on request
Please write to us for further details - services[at]ccamp[dot]res[dot]in