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)

Two types of P-element transformation services:

Injection-only Plan A was closed until further notice (10/31/2021)

Plan A - P-element Basic Service (US$232): 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$290): This service includes injection, generation and identification of transformants (w⁺ or y⁺ included as priced; other markers refer to price table). 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 will *try* to get 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 as of 1x Plan A.

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. 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 occurred, after the unsuccessful 2nd injection, OR 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$405): 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 will *try* to get 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$437): 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.

Three types of PhiC31 transformation services:

Injection-only Plan G was closed until further notice (10/31/2021)

Plan G - PhiC31 Basic Service (US$232): Similar to the above described Plan A. This is an injection-only service and the cost is based on injecting 200 attP G₀ embryos. The G₀ embryos will be collected from the crosses between your selected docking-site strain and the PhiC31-source strain. 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$290): Similar to the above described Plan B and the procedures for Plan G. This service includes injection, generation and identification of transformants (w⁺ or y⁺ included as priced; other markers refer to price table). 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.

When using this service, we will *try* to get at least 1 independent transformants for a reasonably sized construct (smaller than 13 kb) 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$325): 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.

We send you maximum 5 balanced transformants.

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 H+I Service (US$356): 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.

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$464): 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 $11/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 $30 + $12/line.

Plan M - MiMIC Injection Service Package #1 (US$608): This package includes the below 3 items. The original price was >$800, discounted to $608.

When using this service, we will *try* to get at least 2 independent transformants for a reasonably sized construct (smaller than 13 kb) 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.

Plan N - MiMIC Injection Service Package #2 (US$805): This package includes the above Plan M and the below 2 items. The original price was >$1000, now discounted to $805.

4. Service W3 - for Plan M, PCR confirmation of the attP sites for the new MiMIC strain, construct and construct/strain background ($90 includes 1 strain and 1 construct, $69 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$45+24 per additional lines). Must order "Service W3" if you order "Service W4". By default, we only test FOUR lines if we get four or more lines. i.e. the minimum charge is $45 if we only get one line. e.g. If we get 8 lines we only test 4 lines, i.e. $45+$24+$24+$24=$117.

Two types of Genome Engineering CRISPR transformation services:

Plan RH - Genome Engineering Standard Service (US$318): Similar to the above described Plan B and the injection procedures for Plan RG. This service includes injection, generation and identification of CRISPR HDR transformants with visible marker (w⁺ or y⁺ included as priced; other markers refer to price table). We will inject your CRISPR mix into 300 embryos, back-cross SELECTED survival G₀ adults to yw (or suitable stock), expand G₁ transformants by crossing to yw (or suitable stock) again, and ship only the G₂ CRISPR HDR transformants to you. The Plan RH service does NOT include the removal of the Cas9-containing chromosome.

When using this service, we will *try* to get at least ONE independent CRISPR HDR transformants, regardless of the size of the gRNA and donor DNA, due to the high variability of the CRISPR designs. We may re-inject (the second injection) for free only if there is 0 transformant from the first injection round; we will not offer further free injection after the second trial.

Plan RI - Genome Engineering Premium Service (US$354): Similar to the above described Plan C. This service includes injection, generation, identification of transformants (with visible marker) and balancing. We will balance the CRISPR HDR transformants in addition to the Plan RH service. We may directly setup the G₀ crosses using balancers in some situations. The Plan RI service also includes the removal of the Cas9-containing chromosome.

We send you maximum 5 balanced transformants.

We will take the same measures and charge in the same way for the special situations (i.e. toxic or failure transformation) as described in Plan RH.

Plan RH+RI Service (US$386): Plan RI plus an early/partial shipment of Plan RH. i.e. ship Plan RH first (this shipment may not contain all lines), then regular Plan RI shipment when finish.

Two types of All-Other-Injections (i.e. non-P-element, non-PhiC31, non-CRISPR) services:

Injection-only Plan D was closed until further notice (10/31/2021)

Plan D - Other Injections Basic Service (US$232): 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 charge: $114 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$290): Similar to the above described Plan B and the procedures for Plan D. This service includes injection, generation and identification of transformants (w⁺ or y⁺ included as priced; other markers refer to price table).

When using this service, we will *try* to get at least 1 independent transformants . We will send you maximum 5 lines if there are more found in the screening process.

Plan F - Other Injections Premium Service (US$405): 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.

We send you maximum 5 balanced transformants.

Plan E+F Service (US$437): 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.

Plan O Service (US$?): If all of the above standard services may not describe your request. This is the one to let the system assign an order number and the details will be discussed via email. Please ignore the initial money amount showing at the order form.

Molecular Biology services:

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

Service	Price	Description
Z1	US$35	Regular Midiprep. One prep is good for 3x service quantity. We use the Qiagen Midi column.
Z2	US$59	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$81	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$105	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).

Service X1 - Transposable elements mapping for Plan B/C transformants (US$114 per line): When you order "Plan B+X1" or "Plan C+X1", we may 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.

Service X2 - Molecular mapping for Plan RH/RI CRISPR HDR transformants (US$114 per line): When you order "Plan RH+X2" or "Plan RI+X2", we may make genomic DNA extracts of CRISPR HDR transforming flies, perform PCR reactions with primers appropriate for both end of the HDR insertion.

Service W1 - PCR confirmation for PhiC31 Plan H or Plan I transformants (US$34+12 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. No Service W1 for "Own Stock" orders.

Service W2 - PCR confirmation for PhiC31-RMCE transformants (US$45+24 per additional lines): When you order "Service W2", we will make genomic DNA extracts of the transformants flies for the PCR tests and send you the electrophoresis results. No Service W2 for "Own Stock" orders.

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$??? Shipping + US$30 Handling): We only charge a flat rate handling fee per order, no matter how many services are purchased. For instance, if you purchased three "Plan B - Standard", the total will only be US$900. 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$78/108 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. The customer is responsible for all shipping costs including Dead on Arrival.

Special handling fee for flies other than w¹¹¹⁸, yw or listed landing site lines or listed Cas9 lines (US$114): We will charge an extra "Own Stock" handling fee of $114 for the first time you send us the flies. After we finish the project we may keep two bottles for three months for your future orders. If you send next order after that period you may have to pay the new stock fee again. Or you may send us an email to discuss the schedule of the new orders. All "Own Stock" are one-shoot for all Plans.

Screening for adult fluorescence phenotypes (US$59 per Service Quantity): For Plan B/C/H/I/K/M/N/RH/RI/E/F. Screening for adult fluorescent phenotypes (GFP or RFP/DsRed or YFP or CFP), usually on the adult eyes. This can be combined with the w⁺ screening or can be the solo screening of the transformation. Please note if the fluorescence screening need to be done at the larvae stage the additional charge is US$114 per Service Quantity.

Screening for vermilion phenotype (US$59 per Service Quantity): This is usually associated with Plan H/I. The injection can be done at vermilion^- strains and almost all regular vermilion⁺ PhiC31 strains.

Screening for "Loss of Something" phenotype (US$114 per Service Quantity): This has to be ordered with Plan I. When setting up this type of crosses the balancer line(s) has to be used.

Screening extra number of transformants (US$40 per 10 transformants): Screen for more than default number of independent lines (10 for Plan B/C/D/F, 5 for Plan H/I/RH/RI), $35 per 10 additional lines. e.g. if one Plan B generated 35 lines you will be charged (35-10)/10x$40=$100 on top of the regular Plan B price. If the extra lines needs to be balanced, the charge will be additional $12 per line for Plan H/K/RH/RK; $36 per line for Plan B.

PCR confirmation for plasmids (US$34+12 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$34 (first reaction) + $12/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.

Sequencing for gRNA target site / SNP before Plan RG/RH/RI injection (US$114 + US$30 per additional strain): If you wish to check SNP(s) at gRNA target site of the injection strain before ordering Plan RG/RH/RI, we may make genomic DNA extract of the selected strain, perform PCR reaction with primers appropriate for sequencing the gRNA target site. The raw data ONLY, containing the target site sequencing traces of the PCR fragment, will be sent to you by email.

Stock-keeping fee (US$12 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 re-bill 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 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 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 1/1/2024 v4.4