Team:UCL E/Business/Reagents

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Reagents

In order to provide a user-friendly product, the reagents required for use of Darwin Toolbox must be supplied in a ready-to-use format, and be easily accessible to the non-institutional user. On top of these desirable attributes, costs must be kept to a minimum in order to allow the widest possible market to be reached. Therefore, arranging for users to order their reagents as pre-made mastermixes or pre-set kits directly from the Darwin Toolbox website may be the optimal solution to this problem.

An example market for Darwin Toolbox would be iGEM teams, and so quantities and costs have been calculated for such a team. iGEM projects run for approximately 6 months, and a large part of that time is devoted to Biobrick-based cloning. A project kit can be envisaged that encompasses all the enzymes and reagents required (Table 1) and the various clean-up kits required (Table 2).

Component Supplier Cost per unit ($) No. of units required Total cost ($) Taq Polymerase MasterMix NEB 55.00 1 55.00 EcoRI-HF NEB 55.00 1 55.00 XbaI NEB 65.00 1 65.00 SpeI NEB 63.00 2 126.00 PstI NEB 60.00 1 60.00 Quick Ligase kit NEB 97.00 1 97.00 TOTAL 458.00 Table 1. Cloning reagent costs

In addition to this, plasmid miniprep, PCR clean-up, and gel-extraction kits would be required, these are costed in Table 2.

Component Supplier Cost per unit ($) No. of units required Total cost ($) Miniprep kit (100 preps) Anachem Ltd 191.50 1 191.50 Gel extraction kit (100 preps) Anachem Ltd 225.00 1 225.00 PCR clean up (100 preps) Anachem Ltd 225.00 1 225.00 TOTAL 641.50 Table 2. DNA Clean-up kit costs

The total cost of one project kit, buying at retail value would be $1000. However, as many of these items are purchased from the same supplier, and would be ordered in bulk for redistribution to Darwin Toolbox users, the total cost of one project kit could be reduced by as much as X%. Table 3 gives a rough estimate of cost savings associated with bulk orders

TABLE 3

Other markets, such as educational institutes and hobbyists would likely use very similar kits, but most probably require fewer reagents as their usage may not be as frequent as an iGEM team. Therefore, these kits should constantly be reviewed and discussed with the customer community in order to provide the most useful product.

More experienced users, such as field researchers, mould growth investigators, and medical professionals may have very different demands for the Darwin Toolbox. However, these users are less price sensitive than hobbyists and potential iGEM teams, and so bespoke kit design may be a possibility. For instance, providing pre-coated reaction tubes for diagnostic PCR of samples may be a viable optional. An example of this pre-coating is already commercially available for mycoplasma . As entering these markets becomes a more viable option, kit design can be carried out in collaboration with potential users.

Reagent Safety

In order to ensure the safety of our users, Material Safety Data Sheets for the reagents have been compiled, and Standard Operating Procedures and Risk Assessments for using the reagents and Darwin Toolbox have been written.

CU Boulder Collaboration

Reagent Constructs

The CU Boulder iGEM 2013 team have developed a series of constructs and purification methods necessary to produce and isolate restriction enzymes. This would further reduce the cost of reagents for our customers.

Protocols

Experiment design is a key barrier in using our toolbox. Our target markets vary in their previous experience of laboratory work; therefore they require simple proven protocols and troubleshooting advice. The CU Boulder iGEM 2013 team have developed a series of tested protocols as a resource for future iGEM students. We spoke with Phil from their team and discussed using their protocols as a resource for our toolbox users.