Drosophila Facility - User guidelines
Submitting a Drosophila Transgenic/CRISPR Service request:
To submit a request for Drosophila Transgenic services, please click here and register.
After registration, please login and click on "My requests" in the menu bar followed by "New request".
The desired service form can be filled and submitted online.
Registered users can submit a service request by logging in here.
Users are requested to follow the guidelines listed below:
Quantity of DNA:
For P-element mediated transgenesis:
· 40 µg of DNA in ca. 20µl of MilliQ water.
For PhiC31 integrase mediated transgenesis:
- For plasmids:20µg of DNA in ca. 20µl of MilliQ water.
- For BACs/fosmids 4-5µg of DNA in ca. 20µl of MilliQ water.
Quality of DNA:
Please prepare the DNA using a purification procedure such as Qiagen midiprep. The DNA should be free of particulate matter as this will clog microinjection needles. If this proves to be a problem, we will contact you for a fresh sample. Please provide a gel picture of your DNA sample as well as spectrophotometric readings (con. and A260/A280). This can be uploaded in the given fields or can be sent along with the printout of this form and DNA sample.
Information about the DNA:
- Name of plasmid/BAC/fosmid and the selectable marker to be used for scoring transformants.
- Total size of the construct including the insert.
- Any additional information you think might be relevant to this service.
Please fill in the sample submission form online. You MUST take a print out of the completed form for each sample, sign it as well as get it countersigned by PI/lab head and submit this along with the DNA sample. Once this has been received by the facility we will check the form for completeness against the above criteria. Only then the job will be approved/accepted by the facility. Incomplete forms will not be considered.
Terms and Conditions:
- For P-element mediated transgenesis, we offer free re-injection for the constructs (with the same or fresh DNA sample) smaller than 14kb, if we do not get more than 2 independent transformant lines or any transformants from first round of injections.
- For PhiC31 integrase mediated transgenesis, under Site-specific Excel plan we offer free re-injection for the constructs smaller than 14kb (with the same or fresh DNA sample), if we do not get more than 1 or any independent transformant lines from first round of injections.
- For PhiC31 integrase mediated transgenesis, Injections will be performed immediately using the standard strains, whereas for other strains, minimum 6-8 weeks are required for amplifying the stock to the injection scale before injecting the sample. This will be discussed on case-by-case basis.
- BAC/Fosmid will be injected into about 500 embryos of the landing site strain (selected by the user).
- If the DNA construct do not yield more or any transgenics even after second round of injections users may have to pay in full according to the opted service.
- If the user doesn’t get any transformants following our microinjection service, the repeat service will be fully chargeable.
- We do not guarantee the efficiency of designed gRNA, although we use the latest information available to design the gRNA. This will be done in agreement of the user.
- Fly Facility at BLiSc requires that the user has permissions from their Institution’s Internal Biosafety Committee (IBSC) to perform these experiments. Kindly provide a copy of the same.