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Drosophila Embryo Microinjection Service and Pricing

(Please read below, service terms included)

Summary:

Price Description
Plan A US$200 P-element Basic - injection only (ship larvae only)
Plan B US$250 P-element Standard - Plan A plus generation of individual stable transformants (ship transformants)
Plan C US$350 P-element Premium - Plan B plus balancing with balancer chromosome (ship balanced lines)
Plan B+C 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
Plan G US$200 PhiC31 Basic - injection only (ship larvae only)
Plan H US$250 PhiC31 Standard - Plan G plus generation of stable transformants (ship transformants)
Plan I US$280 PhiC31 Premium - Plan H plus balancing (ship balanced lines)
Plan H+I 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 J US$320 PhiC31 Premium Plus Service - Plan I plus removing non-necessary sequences (also ship Plan I flies) (selected strains/vectors only). "Service W2 for Plan J" is recommended
Plan H+J 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 US$400 PhiC31 one-shoot-3x-Plan H-Service for large BACs (BACs >30 kb only). You may choose 1, 2 or 3 strains for the 600+ embryos. Free Service W1 is included
Plan M US$525 MiMIC Injection Service Package #1 - New strain ordering/preparation + Injection + Screening + Balancing
Plan N US$695 MiMIC Injection Service Package #2 - Plan M plus PCR verification of strain/construct (Service W3) and PCR determination of the orientation of transfromants (Service W4)
Plan R US$200 Genome Engineering Basic - injection only (ship larvae only) for CRISPR / TALEN / ZFN
Plan D US$200 Other Injections Basic - injection only (ship larvae only). PiggyBac, Minos, or anything injectable
Plan E US$250 Other Injections Standard - Plan D plus generation of individual stable transformants (ship transformants)
Plan F US$350 Other Injections Premium - Plan E plus balancing with balancer chromosome (ship balanced lines)
Plan E+F US$375 Plan F plus an early/partial shipment of Plan E. i.e. ship Plan E first (this shipment may not contain all lines), then regular Plan F shipment when finish
Service Z1 US$30 Regular plasmid DNA Midiprep. One midiprep is enough for 3 "Service Quantity"
Service Z2 US$50 BAC (P[acman]) Midiprep (<30 kb) plasmids DNA isolation
Service Z3 US$70 BAC (P[acman]) Maxiprep (30-80 kb) plasmids DNA isolation
Service Z4 US$90 BAC (P[acman]) Maxiprep (>80 kb) plasmids DNA isolation
Service X US$100@ Transposable elements mapping for Plan B/C. The final charge will be added when the number of transformant is reported
Service W1 US$30+10@ Genomic PCR confirmation of insertion on landing sites for Plan H or Plan I. The final charge will be added when the number of transformants is reported
Service W2 US$40+20@ Genomic PCR confirmation of insertion on landing sites for Plan J or PhiC31-RMCE. The final charge will be added when the number of transformants is reported. For Plan J, Service W2 includes free Service W1 (see below detailed Plan J & Service W2 descriptions)
Service W3 US$80+60@ For Plan M only. PCR confirmation of the attP sites for the new MiMIC strain, attB construct and construct/strain background. The $80 charge includes 1 strain and 1 construct, additional construct for the same strain costs $60 per construct
Service W4 US$40+20@ Orientation screening for Plan M lines. Must order "Service W3" if you order "Service W4". The final charge will be added when the number of transformants is reported (see below detailed Plan N & Service W4 descriptions)
Fluorescence US$50 Screen for fluorescent phenotypes (GFP or RFP/DsRed or YFP). $50 per service quantity. This can be combined with the w+ screening or can be the solo screening of the transformation
vermilion+ US$50 Screen for vermilion phenotype. $50 per service quantity
New strain US$100 Handling fee of $100 per new strain. You can send us your own strain for injection. Sending it two months prior to the DNA samples recommended
Shipping - You need to provide us your FedEx account. We will use your account for shipping. See below
Handling US$25 Handling fee per order, no mater 1 sample, 10 samples or 100 samples, a fixed $25 handling fee

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 w1118 or 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 the couriers and/or by the 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 or yw, expand G1 transformants by crossing to w1118 or 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 the courier and/or by the 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. toxic or 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. toxic or failure transformation) as described in Plan H.

Plan J - PhiC31 Premium Plus Service (US$320): This service is specific to FlyC31 system (its strains and vectors) and other vectors with the right orientation loxP site. This package includes 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. It's recommended to include the "Service W2 for Plan J" to verify the removal procedures.

Please note we do NOT offer the Plan J service to X-chromosome attP lines (e.g. 24480).

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.

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.


Three special-deal PhiC31 transformation packages:

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 W1 (PCR confirmation for up to 6 lines) is included if there is any transformant. Recommended for testing different landing sites for large BACs. If you wish the resulting lines be balanced, the charge is $10/line (please put this request on the remark); if you wish to receive the regular Plan K shipment plus a later balanced lines shipment, the extra charge is $25 + $10/line.

Plan M - MiMIC Injection Service Package #1 (US$525): This package includes the below 3 items. The original price was $700, discounted to $525.

1. New strain ordering-expanding-preparing ($100 per strain)

This includes the direct BDSC ordering fee (non-refundable if you send us your strain). We order the strain from the BDSC, expand and prepare it for injection. It's recommended to order the Plan N-item#4 (Service W3) below to double check the strain before the injection. We keep the strain for 3 months after the completion of each Plan M.

2. DNA Preparation Service Z1 ($30)

This Plan M package includes the free DNA Preparation Service Z1 (non-refundable if you request to use your DNA directly).

3. Non-standard Plan I - Injection-screening-balancing ($570)

  • setup the injection cage by crossing the MiMIC strain to proper strain with PhiC31 integrase source, collect F1 embryos
  • inject 300+ embryos (i.e. 1.5x quantity of regular Plan H).
  • setup the G0 crosses using proper balancer stocks.
  • screen for "loss of yellow" or "gain of XX" phenotype. We screen up to 8 individual lines.
  • balance the G1 founder using proper balancer stocks
  • The term of "guarantee" is as of Plan H but the number of "1 independent transformant" is changed to "2 independent transformants" due to the fact that the MiMIC integration event is orientational. We will send you maximum 8 lines even if there are more found in the screening process. For constructs smaller than 13kb, we may offer a free 2nd injection if we do not get 2 or more lines from the 1st injection; we may offer a 3rd injection or $120 store credit if we do not get any transformants from the first two round of injections. For constructs larger than 13kb, we may offer a free 2nd injection if we do not get 2 or more lines from the 1st injection; we will charge you full amount regardless of the result of the first two round of transformations.

    In the case of no injection after we prepare the strain, we only charge $100 if we never perform the injection work.

    In the case of one strain multiple constructs injection, we may return $75 per additional constructs for duplicated strain charge. Please contact us in this case.

    Plan N - MiMIC Injection Service Package #2 (US$695): This package includes the above Plan M and the below 2 items. The original price was $880, discounted to $695.

    4. Service W3 - for Plan M, PCR confirmation of the attP sites for the new MiMIC strain, construct and construct/strain background ($80 includes 1 strain and 1 construct, $60 per addition construct). We design the primers, order the oligos, perform genomic DNA extractions and perform all of the PCR works for you. This is very important to verify the strain and the construct before/for the long Plan M progress.

    5. Service W4 - Orientation screening for Plan M lines (US$40 + US$20 per additional lines). Must order "Service W3" if you order "Service W4". By default, we only test 4 lines if we get 4 or more lines. i.e. the minimum charge is $40 if we only get one line. e.g. If we get 8 lines we only test the 4 lines, i.e. $40+$20+$20+$20=$100.


    One package of Genome Engineering (CRISPR / TALEN / ZFN) transformation service:

    Plan R - Genome Engineering 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. Please send us your CRISPR / TALEN / ZFN injection mix (DNAs, RNAs, Oligos, etc) or give us a very clear instruction on how to make the injection mix. You will pay full amount regardless of the number of transformants found in your lab.

    If the injection may contain a visible selection marker, please email us to discuss the possible screening options. We are working very hard to offer mismatch(es) screening services and/or DNA sequencing-based screening services in the near future.

    If you prefer to inject to your own strain, please refer to the below "one-time new strain charge" - $100 per new strain.

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


    Three packages of All-Other-Injections (i.e. non-P-element, non-PhiC31) services:

    Plan D - Other Injections 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, or RNA or any injectable samples from you, then inject them together into 200 embryos. If you prefer to inject to your own strain, please refer to the below "one-time new strain charge", it's $100 per new strain. Usually PiggyBac, Minos, or RNA injections were categorized here.

    The injected larvae will be shipped to you and you are responsible for identifying your own transformants and balancing them (or you are responsible for all of the following experiments). You will pay full amount regardless of the number of transformants found in your lab (or the result of your follow up experiments). There will be no free second injection in ANY situation including shipping mistake by the courier and/or by the customs.

    Plan E - Other Injections Standard Service (US$250): Similar to the above described Plan B and the procedures for Plan D. This service includes injection, generation and identification of transformants.

    The term of "guarantee" is as of plan B but the number of "5 independent transformants" is changed to "1 independent transformant". We will send you maximum 10 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. toxic or failure transformation) as described in Plan B for DNA construct smaller than 13kb. For DNA constructs larger than 13kb, we will charge you full amount regardless of the transformation result.

    Plan F - Other Injections Premium Service (US$350): 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 E service.

    The term of "guarantee" is as of Plan E and we will send you maximum 10 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. toxic or 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 E+F Service (US$375): Plan F plus an early/partial shipment of Plan E. i.e. ship Plan E first (this shipment may not contain all lines), then regular Plan F shipment when finish.


    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 (free of charge).

    A systematic survey of our orders indicated that 62.8% of re-injections using our in-house new DNA prep got significant improvements (i.e. 0 trasnformants to > 5 trasnformants; or 1-2 trasnformants to > 5 trasnformants) v.s. using the original DNA. In the study, we test re-injected our new prep as well as the original DNA side-by-side. The other 37.2% may contain plasmids that may never works (i.e. construct problems, etc.), so the new DNA prep improvement percentage must be even higher. However, we should also take the consideration that the old DNA got frozen at least for one month, i.e. freeze-thaw for at least once.

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

    We will test THREE lines per construct by default (you can request to test more, let us know). Because we cannot predict how many lines will be transformed, we will charge your account after the service. The service report, containing the PCR fragment sequences and the cytosites, will be sent to you by email.

    Service W1 - PCR confirmation for PhiC31 Plan H or Plan I transformants (US$30 + US$10 per additional lines): When you order "Plan H+W1" or "Plan I+W1", we will make genomic DNA extracts of the transformants flies for the PCR tests and send you the electrophoresis results.

    We will test TWO lines per construct by default (you can request to test more, let us know).

    Because we cannot predict how many lines will be transformed, we will charge your account after the service. For example, if you order 1 "Plan H+W1" service for gene-A into the 24483 strain, 1 "Plan I+W1" service for gene-A into the 24487 strain and 1 "Plan I+W1" service for gene-B into the 24488 strain and finally we got 5 w+ flies for each service, we will then only perform PCR for the first 2 w+ lines per service, i.e. 2+2+2=6 y-w+ lines, i.e. $30+$10+$10+$10+$10+$10=$80. The PCR results will be emailed to you after the shipment.

    Service W2 - PCR confirmation for Plan J or PhiC31-RMCE transformants (US$40 + US$20 per additional lines): When you order "Plan J+W2", we will make genomic DNA extracts of the transformants flies for the PCR tests and send you the electrophoresis results. For Plan J, we also include the free "Service W1" to test the corresponding Plan I lines (i.e. totally 2 sets of PCR tests).

    We will test TWO lines per construct by default (you can request to test more, let us know).

    Because we cannot predict how many lines will be transformed, we will charge your account after the service.


    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 (UPS 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/H/I/J/K.

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

    Screening for vermilion phenotype (US$50 per Service Quantity): For Plan H/I. Screening for vermilion phenotype. The injection can be done at vermilion- strains and almost all other regular PhiC31 strains.

    Screening extra number of transformants (US$30 per 10 transformants): Screen for more than default number of independent lines (10 for Plan B/C, 5 for Plan H/I/J), $30 per 10 additional lines. e.g. if one Plan B generated 35 lines you will be charged (35-10)/10x$30=$75 on top of the $250 regular Plan B price. If the extra lines needs to be balanced, the charge will be additional $10 per line.

    PCR confirmation for plasmids (US$30 + US$10 per additional reaction): Quality control restriction enzyme digestions (QC RE check) for verification of new DNA prep made at BestGene is free. However if you also wish to check the PCR of the new DNA prep please send us your primers and the charge is US$30 (first reaction) + $10/additional reaction.

    Sequencing confirmation for plasmids purified by our in-house DNA preps (US$50 + US$20 per additional reaction): Quality control restriction enzyme digestions (QC RE check) for verification of new DNA prep made at BestGene is free. However if your plasmids has potential unstable sequences and you wish to get it sequenced before the injection, we may send out the in-house prepped plasmid for sequencing using your indicated primer(s). This service is non-refundable since it is done by outside providers (actually shipping is the main cost). The charge is US$50 (first reaction) + $20/additional reaction.

    Stock-keeping fee (US$10 per vial per month): We keep backups for 2-4 weeks after the shipping date. If you request to keep any stock beyond this period the extended stock-keeping fee may apply. The minimum charge unit is 1 vial/month (month = calendar month, e.g. 3/4 - 4/3 = one month, 2/27 - 3/27 = two months).

    Returned check charge or rebill fee (US$30): 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 is any additional work 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 deposit 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 12/9/2013 v2.2.4


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