Drosophila Embryo Microinjection Service and Pricing

Summary:

Our staff members have many years of experience in microinjection with different constructs. Our average transformation rate is >15% (transformants/fertile crosses):

200 embryos >125 larvae >100 adults >80 fertile crosses >10 transformants
(>5% if counted as transformants/injected embryos)

Three packages of P-element transformation services:

Plan A - P-element Basic service (US$200): This is an injection-only service and the cost is based on injecting 200 embryos. We obtain DNA sample of the P-element in water from you, then further mix it with the helper plasmid (supplied by you or us) and inject them together into 200 w¹¹¹⁸/yw embryos. The injected larvae will be shipped to you and you are responsible for identifying your own transformants and balancing them. You will pay full amount regardless of the number of transformants found in your lab.

There will be no free second injection in ANY situation including shipping mistake by courier and/or by customs.

Plan B - P-element Standard service (US$250): This service includes injection, generation and identification of transformants. We will inject your DNA sample into 200 embryos, back-cross the survival G₀ adults to w¹¹¹⁸/yw, expand G₁ transformants by crossing to w¹¹¹⁸/yw again, and ship only the G₂ transformants to you. The G₂ transformants will be mailed to you either as adult flies or as larvae.

All injected G₀ adults are individually crossed in single vials! We do not take any short-cut!

When using this service, we guarantee at least 5 independent transformants for a reasonably sized construct (smaller than 13 kb).

In some special situation, if your injected DNA is lethal, we will inform you at once and perform the injection again (free of charge). If the 2nd injection is lethal again, the charge for the "confirmed" lethal injections (200x2 embryo-injections) is US$100.

If your injected DNA has no cloning error, is reasonably sized (smaller than 13 kb) and not lethal but we get no transformant. We will perform the injection again (free of charge) and discuss with you the possible reason of this situation and try to fix the problem. However, if somehow the problem remains, the charge for this situation (200x2 embryo-injections plus crosses for selecting transformants) is usually half of the Plan B charge (i.e. usually US$125. 200% work but only 50% service fee). For easier billing purpose we will send you the invoice of full service amount after the 1st injection and will issue the refund, if occured, after the unsuccessful 2nd injection, as store credit for future orders. If you request the 3rd injection, you will have to pay full amount of the service.

If your injected DNA is larger than 13 kb and we get less than 5 transformants, we will not offer free injection. You are suggested to purchase more service "qty" because usually large constructs are difficult to transform.

Plan C - P-element Premium service (US$350): This is equivalent to "Plan B plus balancing". This service includes injection, identification of transformants and balancing. We will inject your DNA, pick transformants and balance them with balancer chromosomes. We use single balancers. This service takes longer time and you will be charged the full amount regardless of which chromosome(s) the P-elements are incorporated in.

Similar to the description of Plan B above, we guarantee at least 4 independent transformants for a reasonable sized construct (smaller than 13 kb).

We will take the same measure and charge in the same way for the special situations (i.e. failure transformation) as described in Plan B.

Plan B+C service (US$375): Plan C plus an early/partial shipment of Plan B. i.e. ship Plan B first (this shipment may not contain all lines), then regular Plan C shipment when finish.

Four packages of PhiC31 transformation services:

Plan G - PhiC31 Basic service (US$200): Similar to the above described Plan A. This is an injection-only service and the cost is based on injecting 200 attP embryos. For P[acman] docking sites injections, some of the G₀ embryos will be collected from the crosses between your selected docking-site strain and the PhiC31-source strain. For some P[acman] docking site and all FlyC31 landing site inejctions, the G₀ embryos will be collected directly because these strains already contain the PhiC31 source. The injected G₀ larvae will be shipped to you and you are responsible for identifying your own transformants and remove the PhiC31 source (by phenotype). You will pay full amount regardless of the number of transformants found in your lab.

There will be no free second injection in ANY situation including shipping mistake by courier and/or by customs.

Plan H - PhiC31 Standard service (US$250): Similar to the above described Plan B and the procedures for Plan G. This service includes injection, generation and identification of transformants. We will inject your attB DNA sample into 200 embryos, back-cross SELECTED survival G₀ adults to yw, expand G₁ transformants by crossing to yw again, and ship only the G₂ transformants to you.

The term of "guarantee" is as of plan B but the number of "5 independent transformants" is changed to "1 independent transformant" due to the fact that in most cases you only need one line to perform your experiment. We will send you maximum 5 lines if there are more found in the screening process.

We will take the same measure and charge in the same way for the special situations (i.e. failure transformation) as described in Plan B for construct smaller than 13kb. For constructs larger than 13kb, we will charge you full amount regardless of the transformation result.

Plan I - PhiC31 Premium service (US$280): Similar to the above described Plan C. This service includes injection, generation, identification of transformants and balancing. We will balance the transformants in addition to the Plan H service.

The term of "guarantee" is as of Plan H and we will send you maximum 5 balanced transformants if there are more found in the screening process.

We will take the same measure and charge in the same way for the special situations (i.e. failure transformation) as described in Plan H.

Plan J - FlyC31 Specific service (US$320): This service is specific to FlyC31 system (its strains and vectors), including injection, generation, identification of transformants, balancing and eliminating non-necessary sequences. We will provide the stable lines containing your transgene by Cre-mediated excision leaving ONLY one loxP site and one attL site. We will also send you the corresponding Plan I balanced flies.

We will take the same measure and charge in the same way for the special situations (i.e. failure transformation) as described in Plan H.

Plan H+I service (US$305): Plan I plus an early/partial shipment of Plan H. i.e. ship Plan H first (this shipment may not contain all lines), then regular Plan I shipment when finish.

Plan H+J service (US$345): Plan J plus an early/partial shipment of Plan H. i.e. ship Plan H first (this shipment may not contain all lines), then regular Plan J shipment when finish.

Plan K - PhiC31 non-Standard service - PhiC31 one-shoot-3x-Plan H-Service for large BACs of >30 kb only (US$400): Similar idea of 1.5x price of Plan H and we inject 600+ embryos at once, no free re-injection and no store credits if no transformants. You may choose 1, 2 or 3 strains for the 600+ embryos. Free Service W (PCR confirmation for up to 6 lines) is included if there is any transformants. Recommended for testing different landing sites for large BACs.

Other services:

Service Z - DNA preparation Services: We also provide additional DNA preparation assistance and the costs are as follow:

Service	Price	Description
Z1	US$30	Regular Midiprep. One prep is good for 3x service quantity. We use the Qiagen Midi column.
Z2	US$50	BAC Midiprep for constructs smaller than 30 kb. For P[acman] constructs requires the EPI300 cells and induction before the DNA prep. We use the Qiagen Midi column.
Z3	US$70	BAC Maxiprep for constructs between 30-80 kb. For P[acman] constructs requires the EPI300 cells and induction before the DNA prep. We use the Qiagen Large Construct Kit.
Z4	US$90	BAC Maxiprep for constructs larger than 80 kb. For P[acman] constructs requires electroporation to the EPI300 cells and induction before the DNA prep. We use the Qiagen Large Construct Kit.

We highly recommend you to include the proper plasmid map along with your sample so that we can do a quality control digest after the DNA prep.

Service X - Inverse PCR for P-element transformants (US$100 per line): When you order "Plan B+X" or "Plan C+X", we will make genomic DNA extracts of the P-element transforming flies to be digested with proper restriction enzyme, followed by ligation and PCR reactions with primers appropriate for the end of P-element.

Service W - PCR confirmation for PhiC31 Plan H/I/J transformants (US$30 + US$10 per additional lines): When you order "Plan H+W", "Plan I+W" or "Plan J+W", we will make genomic DNA extracts of the w⁺ flies for the PCR tests and send you the electrophoresis results. For "Plan J+W" we will also include the test for the "Plan I+W" files and the charge will be US$40+US$20 per additional lines.

Other charges:

Shipping and Handling (US$25 handling): We only charge a US$25 flat rate handling fee per order, no matter how many services are purchased. For instance, if you purchased three of the "Plan B - Standard", the total will only be US$775.00. We will use your FedEx account for sending back flies, so please provide the number when you are registering or ordering. It will be US$50/100 EXTRA shipping charge for domestic/international (including Canada) customers if you do not provide us your FedEx number (DHL or other couriers are fine, please email us for details). In some rare cases, if re-ship is required/requested, we will use your FedEx account; if you did not include any courier account, we will charge you the shipping fee again.

Special handling fee for flies other than w¹¹¹⁸/yw or listed landing site lines (US$100): We will charge an extra handling fee of $100 for the first time you send us the flies. After we finish the project we will keep two bottles for three months for your future orders. If you send next order after that period you will have to pay the $100 again. Or you can send us an email to discuss the schedule for new orders. Special strain does not qualify for the "guarantee" mentioned above for Plan B/C/E/F/H/I.

Screening fluorescence phenotypes (US$50 per Service Quantity):For Plan B/C/H/I/J. Screening of fluorescent phenotypes (GFP or RFP). This can be combined with the w⁺ screening or can be the solo screening of the transformation.

Screening extra number of transformants (US$25 per 10 transformants): Screen for more than default number of independent lines (10 for Plan B/C, 5 for Plan H/I/J), $25 per 10 additional lines. e.g. if one Plan B generated 35 lines you will be charged (35-10)/10x$25=$50 on top of the $250 regular Plan B price. If the extra lines needs to be balanced, the charge will be $100 per 10 Plan B lines; $30 per 10 Plan I lines. In short, $2.50 per extra Plan B lines; $12.50 per extra Plan C lines.

Returned check charge or rebill fee: US$30 will be charged for extra bank/courier administration fee if we got returned check, less wire amount or invalid courier account related to your payment/shipment.

Other services fees: There are no hidden fees! All fees are listed above. However we will inform you in advance if there are any additional works needed to be performed.

Current Promotions:

Pre-pay orders (10% off): For all customers charge their account prior to their ordering. Providing a PO number does not qualify this discount. This discount needs the fund in our account before the actual ordering. There is a minimum of US$2,000.00 per prepay transaction.

Immediate payment orders (2% off): For all orders (online or offline) submitted together with credit card information. Payment has to be received along with your order form and DNA sample(s).

updated 10/24/2011 v2.1.2