Biohacking Costing and Prices

• Autoclave strips. These supposedly tell you if the autoclaving has been done at the required conditions, unlike autoclave tape which only tells you if the temperature has been reached.
• If extractions keep failing, lets look at some extraction kits. This one recommended by Tom is ∼£100 for 100 reactions or ∼£250 for 500 extractions.
• A more consistent and effective way of crushing/homogenising small amounts of tissue for DNA extraction. Blenders are good but not for small quantities. Pestles are ok, but not great. One idea is to make a small pestle by melting a pipette tip then using a PCR tube as a mortar. Some labs use liquid nitrogen or a sonicator.
• Spectrophotometer for measuring DNA concentration. Simon and Tom are on this.
• Good microscope enabling us to see cells (Possible mag: 100x, 400x, 1000x).
• Better imaging system

Costs (assuming prices on wet stuff)

Per Chelex DNA extraction:

• Chelex: £0.33 (0.15g)
• Pasteur pipettes: £0.06 (1 disposable)
• Tubes: £0.04 (1 for chelex incubation, 1 to save supernatant)
• Tips: £0.06 (1 for mixing chelex, 1 removing supernatant)

Total per sample: £0.49

Per PCR reaction Assume 25ul total volume. 12.5ul Taq Readymix, 5ul primers

• Taq Readymix: £0.72 (12.5ul)
• Primers: Not calculated but likely to be small
• Tubes: £0.02 (1 tube for PCR)
• Tips: £0.15 (Assuming 1 tip for template, 1 for PF, 1 for PR, 1 for dH20, 1 for Taq)
• Paraffin: Negligible

Total per sample: £0.89 + primers

Per restriction digest for blood typing: This estimate may be wrong. Need to calculate quantities definitively

• KPN1 and ALU1: £2.08

Total per sample: £2.08 + another gel

Per sample on a gel: For each sample:

• Tubes: £0.02 (1 to mix loading buffer with PCR product)
• Tips: £0.03 (1 to add product to loading buffer and then load into well)
• Loading buffer: Negligible

Total per sample: £0.05

Per gel: Assume a 1% 50ml agarose gel with 100ml buffer and 2.2ul EtBr

• Agarose: £0.23
• TBE: £0.13
• Ladder: £0.30
• Tips: £0.03 (1 to add loading buffer to all tubes)
• dH20: Negligible
• EtBr: Negligible

Total per gel: £0.69

So for a chelex extraction, PCR and gel it costs at least £1.43 per sample. A gel costs £0.69, so a typical session involving 4 extractions + PCRs, followed by a gel costs at least £6.14.

Ideas for cost+efficiency savings

• Cheaper Taq would make the biggest contribution. Investigate NEB (won't sell to us at the moment), web scientific and nbs.
• Smaller gels and gel box could halve amount of TBE and agarose used.
• Reduce concentration of agarose and TBE in gel and buffer. E.g. we could use 0.5% TBE not 1%.
• Use TAE instead of TBE (TAE is 1/2 to 1/3 the price of TBE). However it is claimed TBE is better for small DNA lengths like ours
• Buy powdered TBE. NBSBio - £33.60 for 1 litre of 10x (50p per gel+buffer), and £76.80 for 5L (23p per gel+buffer). Dry premix powders are even better value than this. £18 for 1L (27p per gel+buffer), and £28 for 4L (13p per gel+buffer).
• Make our own TBE. Recipe here. Advantage that we could also make TE.
• Invitrogen do http://products.invitrogen.com/ivgn/product/15581044# cheaper TBE] but won't sell to us atm
• Cheaper sources for tubes and tips e.g. ebay
• Heated lid making paraffin unnecessary. Save on paraffin + extra tubes for mixing loading buffer with PCR product as loading buffer could be added directly to it if there are no further plans for it. Mostly efficiency saving rather than cost.
• Same result could be achieved by using a taq readymix with loading buffer included (e.g. sold by webscientific)
• Larger quantities of agarose. Our agarose from NBSBio works out at £0.23 per gel for 100g, but if we bought 500g it would be £0.18 per gel. Not a massive saving though.

DIY Biology group based at London Hackspace, main wiki page here.

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