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Fly Extract


“Fly Extract” from OregonR adults is required as an additive to culture many insect cell lines (e.g. Drosophila OSS, OSC, fGS/OSS, CME Cl.8+, and other lines derived from ovaries or imaginal discs). The VDRC has been providing Fly Extract for a number of Vienna Biocenter groups and now extends this service to the Drosophila community.

The VDRC Fly Extract is produced using a protocol developed by the lab of Julius Brennecke (IMBA, Vienna), which has been successfully growing OSC cells using this extract since 2010 (Sienski et al., Cell 2012).

We recommend using this extract at the following concentrations:

VDRC Fly Extract
aliquot volume		 Dilute into
(medium volume)		 Final concentration
VDRC Fly Extract*		 Cell lines
1.25ml 100ml 1.25% Wing discs (e.g. Cl.8R; CME W1)
Leg discs (e.g. CME L1; L127D6)
5ml 100ml 5% Ovary (e.g. OSC; OSS; fGS/OSS)

* Please note, the VDRC Fly Extract is 2x more concentrated than that available from the DGRC, so only half the volume and half the final concentration are required for the equivalent functionality.

  1. First check that your institute/shipping address is able to receive packages containing dry ice.
  2. Download the Fly Extract order form ( academic users / company users).
    Please note: Unlike the fly stocks and DNA constructs, it is currently not possible to place an order for VDRC Fly Extract online. Only orders submitted via the VDRC Fly Extract order form will be accepted.
  3. Complete all required fields, print out, add your signature, scan and return the order form by email to office@vdrc.at.
  4. Once we receive your order form, we will send an invoice to the contact email address and billing email address, as you have entered them on your order form.
  5. You should pay the invoice upon receipt by wire transfer. It is not possible to pay the invoice by credit card or by cheque.
  6. Your order will be prepared as soon as we are informed by our accounting department that the payment has been received.
  7. Your order will be shipped in packages containing dry ice by the courier specified in your order form.
  8. You will receive an email with the shipping details and the courier tracking number.
  9. You should receive the Fly Extract in a frozen condition.
  10. Currently each new order of Fly Extract will require the form to be filled in again.

Please note: Due to the different temperature and legal documentation requirements, orders of fly stocks/DNA constructs and Fly Extract will be treated as completely independent orders and cannot be sent in a combined shipment.

Academic/non-profit:
Item ID Item description Unit size Price/unit (not including tax)
VFE1.25 VDRC Fly Extract, 1.25ml aliquot 1.25ml 9,50 Eur
VFE5 VDRC Fly Extract, 5ml aliquot 5ml 38 Eur

Companies:
Item ID Item description Unit size Price/unit (not including tax)
VFE1.25 VDRC Fly Extract, 1.25ml aliquot 1.25ml 12 Eur
VFE5 VDRC Fly Extract, 5ml aliquot 5ml 48 Eur

An additional flat fee of 15 Eur per order will be added for packaging and dry ice.

Please note, the VDRC Fly Extract is 2x more concentrated than that available from the DGRC, so only half the volume and half the final concentration are required for the equivalent functionality. For further information about volumes and concentrations please see the section: "How to prepare media using VDRC Fly Extract".

Fly Extract should be frozen at -20°C on arrival. It is still good after 6 months storage at -20°C (and it will most likely keep for longer but this has not been tested yet).

Thawing and refreezing should be avoided if possible, but one thaw and refreeze leads to no obvious loss of quality in our hands.

The concentration of Fly Extract required depends on the cell line. When using VDRC Fly Extract we recommend the following:


Media for ovary cell lines (OSS, OSC and fGS/OSS)

(M3 + 10%FBS + 10µg/ml insulin + 60µg/ml glutathione + 5% VDRC Fly Extract)


Components

  • Shields and Sang M3 Insect media (Sigma-Aldrich, S3652-500ML).
  • Fetal Bovine Serum (FBS) (e.g. Sigma, F2442-50ML). Heat inactivate at 60°C for 30 minutes. Store in 10ml aliquots.
  • Insulin: (Sigma-Aldrich: I6634-100MG). Make 1mg/ml (=100x) stock: 100mg in 100ml Milli-Q H2O + 8.5ul of 37% HCl and store in 1ml aliquots.
  • L-Glutathione reduced: (Sigma-Aldrich: G6013-25G). Dilute 6g/100ml Milli-Q H2O and store in 1ml aliquots.
  • VDRC Fly Extract (FE): 5ml aliquot.

Prepare 100ml Medium:

  • 83ml M3 Media
  • 10ml FBS
  • 1ml Insulin
  • 1ml Glutathione
  • 5ml VDRC Fly Extract

Mix all components and filter sterilize through a 0.2uM sterile filter. Store at 4°C.


Media for Cl.8R cells

(M3 + 2% FCS + 5 µg/ml insulin + 1.25% VDRC Fly Extract)


Components

  • Shields and Sang M3 Insect media (Sigma-Aldrich, S3652-500ML).
  • Fetal Bovine Serum (FBS) (e.g. Sigma, F2442-50ML). Heat inactivate at 60°C for 30 minutes. Store in aliquots.
  • Insulin: (Sigma-Aldrich: I6634-100MG). Make 1mg/ml (=100x) stock: 100mg in 100ml Milli-Q H2O + 8.5ul of 37% HCl and store in aliquots.
  • VDRC Fly Extract (FE): 1.25ml aliquot.

Prepare 100ml Medium:

  • 93ml M3 Media
  • 5ml FBS
  • 0.5ml Insulin
  • 1.25ml VDRC Fly Extract

Mix all components and filter sterilize through a 0.2uM sterile filter. Store at 4°C.

Continuation of funding for VDRC by the City of Vienna and the Austrian Ministry of Science is dependent on documented evidence of contributions to scientific output. Therefore, we request you to acknowledge the VDRC when you obtain resources from us.

For Fly Extract, a simple statement is sufficient and can either be placed in the Materials and Methods section or in the Acknowledgements.


Suggested format:

Fly Extract was obtained from the Vienna Drosophila Resource Center (VDRC, www.vdrc.at).