Every consumable, instrument, reagent, and tip needed to take a buccal swab through to a basecalled BAM. Costs are per-unit where I had access to an open pack and per-pack where I didn't. Assumes 2 dry runs (water only, to practice the protocol) plus 1 real run.
Hardware, single-use plastics, and shared lab kit.
| Item | Why | Cost | Units |
|---|---|---|---|
| MinION Mk1D Sequencer | Drives the trans-membrane voltage on the flow cell and records the current changes as DNA passes through each pore. | ~$3,200 | 1 |
R10.4.1 Flow Cell FLO-MIN114 |
Consumable holding the nanopore array; ~1,200 active pores at start. | ~$900 | 1 |
Flow Cell Wash Kit EXP-WSH004 |
Nuclease wash to recover blocked pores mid-run; enables a second library load. | ~$17/wash | 1–2 washes |
| Microcentrifuge | Column-based DNA extraction spins; also needed for Covaris g-TUBE shearing. | ~$65 | 1 |
| Heat block / dry bath | 56 °C FFPE repair, 65 °C end-prep, room-temp ligation. No heated lid — watch for condensation on tube lids at 65 °C. | ~$65 | 1 |
| Micropipettes (P20, P200, P1000) | Liquid handling throughout. See note below on accuracy. | ~$25 borrowed / ~$250–500 refurbished | 3 pipettes |
| Tips, 10 µL (box of 96) | Small-volume pipetting (2–10 µL enzyme additions, residual removal). | ~$7–13 | 1 box (40 used) |
| Tips, 200 µL (box of 96) | Mid-volume pipetting (DNA transfers, bead cleanups, washes). | ~$7–13 | 1 box (85 used) |
| Tips, 1000 µL (box of 96) | Large-volume pipetting (LFB washes, flow cell priming, extraction washes). | ~$7–13 | 1 box (28 used) |
| Wide-bore 200 µL tips | Handling HMW gDNA without shearing (pre-Covaris transfers). | ~$13–20 | ~5 tips |
| Buccal swab kit | Sample collection from cheek cells. | ~$20–32 | 4 swabs |
Monarch Spin gDNA Extraction Kit T3010S |
Column-based gDNA extraction from buccal swabs. | ~$2.30–3.10/prep | 4 preps |
| Nuclease-free water | Elution, dilution, and dry-run practice. | ~$13 | 1 bottle (~50 mL) |
| DNA LoBind tubes, 1.5 mL | Low-bind tubes to prevent DNA loss on tube walls. | ~$0.09–0.14/tube | ~40–50 |
Minimum set is P20 + P200 + P1000 (three pipettes). The smallest volume in the ligation protocol is ~2.5 µL for enzyme additions, within the P20's accurate range (2–20 µL). A P10 (1–10 µL) is nice-to-have for small-volume work but not required.
| Phase | 10 µL | 200 µL | 1000 µL | Total |
|---|---|---|---|---|
| Library prep (×3 runs) | 27 | 48 | 12 | 87 |
| DNA extraction (4 swabs, real only) | 4 | 17 | 8 | 29 |
| Covaris shearing (real only) | 0 | 2 wb | 0 | 2 |
| Flow cell priming + loading (real only) | 2 | 4 | 3 | 9 |
| Subtotal | 33 | 71 | 23 | 127 |
| Spares (~20%) | 7 | 14 | 5 | 26 |
| Total | 40 | 85 | 28 | 153 |
One box of 96 per tip size covers it. Library prep and flow cell counts are exact per the ONT protocol (HVSW_9217_v114_revB); DNA extraction is estimated at ~7 tips per swab from the Monarch kit.
SQK-LSK114The reagent block. Three of these line items are bulk packs where you only use one or a few units; see the bulk-pack table further down.
| Item | Why | Cost | Units |
|---|---|---|---|
| SQK-LSK114 (Ligation Sequencing Kit V14, 6 rxns) | End-repair, dA-tailing, T4 ligase adapter ligation; includes AMPure XP beads, buffers, and adapters. | ~$599 | 1 rxn (kit has 6) |
NEBNext Companion Module v2 E7672S (24 rxns) |
FFPE Repair v2, Ultra II End-prep, Salt-T4 DNA Ligase, Thermolabile Proteinase K. All NEB reagents needed for SQK-LSK114 in one purchase. | ~$54/rxn | 1 rxn |
| Covaris g-TUBE | Controlled DNA shearing to ~6–8 kb N50 via centrifugation; produces the narrow fragment distribution needed for optimal adaptive sampling. | ~$60–72/tube | 1 tube |
| Magnetic rack (DIY) | Pulls AMPure beads to the side of the tube during the two cleanup steps. | ~$13–20 | 1 rack |
| N52 neodymium block magnets (10×5×2 mm) | For the DIY rack; two stacked per tube slot. | ~$0.66 each | 10–15 |
| 0.2 mL PCR tubes | For enzyme incubation steps. | ~$7–13 (pack of 100) | ~10 tubes |
| 80% ethanol (fresh, molecular grade) | Bead wash steps (two washes per cleanup, two cleanups per prep). | ~$13–20 | ~20 mL (real run only) |
2 dry runs + 1 real run.
| Protocol step | Tubes per run | Runs | Total tubes |
|---|---|---|---|
| DNA extraction (4 preps) | ~8 | 1 (real only) | ~8 |
| Library prep (ligation) | ~6 | 3 | ~18 |
| Flow cell priming + loading | ~2 | 1 (real only) | ~2 |
| Spares / mistakes | — | — | ~5–10 |
| Total | ~37 |
| Item | Cost (per-unit) | Cost (full pack) | Notes |
|---|---|---|---|
| SQK-LSK114 | ~$100 | ~$599 | 6 rxns per kit, you use 1. |
NEB reagents E7672S | ~$54 | ~$1,275 | 24 rxns per kit; per-unit assumes access to an open pack at a lab. |
| Covaris g-TUBE | ~$72 | ~$710 | 10 per pack; per-unit assumes access to an open pack. |
| Magnetic rack (DIY) | ~$13 | ~$13 | |
| Other consumables | ~$25 | ~$25 | PCR tubes, ethanol. |
| Total | ~$264 | ~$2,622 |
Items sold in packs where you only need one or a few units. This is the gap home sequencing has to bridge somehow — either through bulk-buying schemes, shared access at an institution, or per-rxn rentals.
| Item | Pack size | Pack price | Unit price | Units needed | Your cost (unit) | Your cost (pack) |
|---|---|---|---|---|---|---|
| SQK-LSK114 (Ligation Kit) | 6 reactions | ~$599 | ~$100/rxn | 1 | ~$100 | ~$599 |
NEBNext Companion Module v2 E7672S |
24 reactions | ~$1,275 | ~$54/rxn | 1 | ~$54 | ~$1,275 |
Covaris g-TUBE 520079 |
10 tubes | ~$710 | ~$72/tube | 1 | ~$72 | ~$710 |
Monarch gDNA Extraction Kit T3010S |
50 preps | ~$115–152 | ~$2.30–3.10/prep | 4 | ~$9–13 | ~$115–152 |
| DNA LoBind tubes, 1.5 mL | 250 tubes | ~$23–35 | ~$0.09–0.14/tube | 25–40 | ~$3–5 | ~$23–35 |
Flow Cell Wash Kit EXP-WSH004 |
6 washes | ~$95 | ~$17/wash | 1–2 | ~$17–32 | ~$95 |
| 0.2 mL PCR tubes | 100 tubes | ~$7–13 | ~$0.06–0.13/tube | ~10 | ~$0.65–1.30 | ~$7–13 |
Every reagent required for DNA repair, end-prep (protocol § 4) and adapter ligation (protocol § 5), and which kit it ships in. Reference: HVSW_9217_v114_revB.
| Reagent | Protocol step | Ships in |
|---|---|---|
| NEBNext FFPE DNA Repair Buffer v2 | 4.3 (FFPE repair) | NEB Companion E7672S |
| NEBNext FFPE DNA Repair Mix v2 | 4.3 (FFPE repair) | NEB Companion E7672S |
| Thermolabile Proteinase K | 4.7 (post-repair) | NEB Companion E7672S |
| NEBNext Ultra II End Prep Enzyme Mix | 4.11 (end-prep) | NEB Companion E7672S |
| AMPure XP Beads (AXP) | 4.16, 5.9 (cleanups) | SQK-LSK114 |
| Ligation Buffer (LNB) | 5.5 (ligation) | SQK-LSK114 |
| Salt-T4 DNA Ligase | 5.5 (ligation) | NEB Companion E7672S |
| Ligation Adapter (LA) | 5.5 (ligation) | SQK-LSK114 |
| Long Fragment Buffer (LFB) | 5.12, 5.13 (washes) | SQK-LSK114 |
| Elution Buffer (EB) | 5.15 (library elute) | SQK-LSK114 |
| Nuclease-free water | 4.2, 4.23 (volume/elute) | supplied separately |
| Freshly prepared 80% ethanol | 4.20, 4.21 (bead washes) | supplied separately |
With SQK-LSK114 + NEBNext Companion Module v2 in hand, the only extras to source are nuclease-free water and molecular-grade ethanol.
Fresh tip per reagent addition; tip reuse only for pipette-mixing within the same tube. Referenced against ONT protocol HVSW_9217_v114_revB.
| Step | Action | Volume | Tip size |
|---|---|---|---|
| 4.2 | Transfer gDNA to PCR tube | 46 µL | 200 µL (wide-bore) |
| 4.2 | Add NFW to adjust volume | varies | 20 µL |
| 4.3 | Add FFPE Repair Buffer v2 | 7 µL | 10 µL |
| 4.3 | Add FFPE Repair Mix v2 | 2 µL | 10 µL |
| 4.7 | Add Thermolabile Proteinase K | 2 µL | 10 µL |
| 4.11 | Add End Prep Enzyme Mix | 3 µL | 10 µL |
| 4.15 | Transfer to LoBind tube | 60 µL | 200 µL |
| 4.16 | Add AMPure XP beads | 60 µL | 200 µL |
| 4.19 | Remove supernatant | ~120 µL | 200 µL |
| 4.20 | Add ethanol wash 1 | 200 µL | 200 µL |
| 4.20 | Remove ethanol wash 1 | 200 µL | 200 µL |
| 4.21 | Add ethanol wash 2 | 200 µL | 200 µL |
| 4.21 | Remove ethanol wash 2 | 200 µL | 200 µL |
| 4.22 | Remove residual ethanol | ~5 µL | 10 µL |
| 4.23 | Add NFW to resuspend pellet | 61 µL | 200 µL |
| 4.25 | Remove eluate | 61 µL | 200 µL |
| Step | Action | Volume | Tip size |
|---|---|---|---|
| 5.5 | Add Ligation Buffer (LNB) | 25 µL | 200 µL |
| 5.5 | Add Salt-T4 Ligase | 10 µL | 10 µL |
| 5.5 | Add Ligation Adapter (LA) | 5 µL | 10 µL |
| 5.6 | Mix reaction | 100 µL | 200 µL |
| 5.9 | Add AMPure XP beads | 40 µL | 200 µL |
| 5.11 | Remove supernatant | ~140 µL | 200 µL |
| 5.12 | Add LFB wash 1 | 250 µL | 1000 µL |
| 5.12 | Remove LFB wash 1 | 250 µL | 1000 µL |
| 5.13 | Add LFB wash 2 | 250 µL | 1000 µL |
| 5.13 | Remove LFB wash 2 | 250 µL | 1000 µL |
| 5.14 | Remove residual supernatant | ~5 µL | 10 µL |
| 5.15 | Add Elution Buffer | 33 µL | 200 µL |
| 5.17 | Remove eluate | 33 µL | 200 µL |
| Step | Action | Volume | Tip size |
|---|---|---|---|
| 6.2 | Pipette FCT into priming mix | 30 µL | 200 µL |
| 6.2 | Pipette FCF into priming mix | 1170 µL | 1000 µL |
| 6.6 | Draw back buffer from flow cell | ~20–30 µL | 1000 µL |
| 6.7 | Load priming mix (first flush) | 500 µL | 1000 µL |
| 6.9 | Pipette Sequencing Buffer (SB) | 100 µL | 200 µL |
| 6.9 | Pipette Library Beads (LIB) | 68 µL | 200 µL |
| 6.9 | Pipette DNA library | 32 µL | 200 µL |
| 6.10 | Load priming mix (second flush) | 500 µL | 1000 µL |
| 6.12 | Load library into flow cell | 200 µL | 200 µL |