Drosophila Embryo Microinjection Service and Pricing

Summary:

Three packages of P-element transformation services:

Our staff members have many years of experience in microinjection with different P-element constructs. Our average transformation rate is 13-17% (transformants/fertile crosses):

200 embryos 125 larvae 80 adults 60 fertile crosses 10+ transformants
(about 4-5% if counted as transformants/injected embryos)

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 to 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 adult to w¹¹¹⁸/yw crosses are individual male-female done 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 US$125 (200% work but only 50% service fee). 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 you 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.

Three packages of Cre/loxP Recombination-Mediated Cassette Exchange (RMCE) transformation services:

Plan D - Cre/loxP RMCE 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 embryos. We obtain DNA sample of the RMCE donor construct in water from you, then further mix it with the helper plasmid (supplied by us, expressing Cre recombinase) and inject them together into 200 of your specified landing site embryos. The injected larvae will be shipped to you and you are responsible for identifying your own transformants. 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 E - Cre/loxP RMCE Standard service (US$250): Similar to the above described Plan B. This service includes injection, generation and identification of transformants. We will inject your RMCE donor sample to 200 of your specified landing site embryos, back-cross the survival G₀ adults to yw, expand G₁ transformants (y^-w⁺) by crossing to yw again, and ship only the G₂ transformants to you.

There is no "guarantee" for plan E due to the fact that RMCE system has very low transformation efficiency. However, you will still receieve ONE free re-injection if the first trial yield no w⁺. We will send you maximum 5 transformants if there are more found in the screening process.

We will charge you full amount for plan E regardless of the transformation result.

Plan F - Cre/loxP RMCE Premium service (US$280): Similar to the above described Plan C. This service includes injection, generation, identification of transformants and balancing. We will inject your Cre/loxP RMCE donor construct, pick transformants and balance them with balancer chromosome.

The term of "guarantee" is as of plan E and we will send you maximum 5 balanced transformants if there are more found in the screening process. For failure transformations (i.e. two unsuccessful injections), we will only charge you the amount for Plan E and will not charge the amount for balancing.

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 100-200 attP embryos. For P[acman] docking sites injections, the G₀ embryos will be collected from the crosses between your selected docking-site strain and the PhiC31-source strain. For 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 to 100-200 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 terms 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.

The term of "guarantee" is as of Plan H and we will send you maximum 5 stable lines and their corresponding balanced ones 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.

Other services:

Service Z - Additional DNA preparation (US$30): We also provide additional DNA preparation assistance and the cost is US$30 per DNA sample. The rest of the service is the same as described. 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 Y - PCR confirmation for RMCE transformants (US$30 + US$10 per additional lines): When you order "Plan E+Y" or "Plan E+Z+Y", we will make genomic DNA extracts of the w⁺ flies and perform PCRs. The PCRs are using primers pairing to the left end and the right end of the cassette: the left end gives ~800 bp product, while the right end yields longer one depends on your insert. We will perform a long PCR reaction for the right end but we can not guarantee it will work if your insert is too big. We will do maximum 2 lines per order by default (you can request to do more, let us know!). For example, if you order 1 service for gene-A into the A10 strain, 1 service for gene-A into the A11 strain and 1 service for gene-B into the A10 strain and finally we got 3 y^-w⁺ flies for each service, we will then only perform PCR for the first 2 y^-w⁺ lines per service, i.e. 2+2+2=6 y^-w⁺ lines, i.e. $30+$10+$10+$10+$10+$10=$80.

Because we can not predict how many lines will be transformed, we will charge your account when the number of y^-w⁺ is reported. The PCR results will be sent together with your fly shipment or emailed to you before the shipment per your request.

Service X - Inverse PCR for P-element transformants: To be announced

Service W - PCR confirmation for FlyC31 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 with your plasmid constructs as the insert size control 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 as a 2nd control 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: 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.

Other services fees: There are no hidden fees! All fees are listed above. However we will inform you if there are any additional service(s) needed to be performed. For example, if there is a need to sequence the construct to make sure the correctness of the clone we will contact you and after your confirmation we will charge you $20 and perform the experiment.

Current Promotions:

Pre-pay orders (10% off): For all customers charge their account prior to their ordering.

Immediate payment orders (5% 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).

Providing a PO number does not qualify these discounts. These discounts need the fund in our account before the actual ordering.

Referral bonus (5%): Refer to a friend and you will earn 5% from his second order onward. Referral bonus can only be used to purchase service at our company and no cash value.

Citation Bonus (one free plan B/H): Please acknowledge BestGene Inc in the resultant publication and forward us the PDF file. We will credit one plan B/H for your compliment. Citation bonus can only be used to purchase service at our company and no cash value.

updated 10/08/2008 v1.2.4