| TaqMan | TaqMan Endpoint | HRM | AS-PCR | SSOP-ELISA | PCR Dot Blot | HOLA |
---|---|---|---|---|---|---|---|
Specialist equipment required | Real-time PCR machine | PCR thermocyler Fluorimeter | High-spec real time PCR machine | PCR thermocycler Gel electrophoresis and imaging equipment | PCR thermocycler Shaking incubator Multichannel pipette *optional ELISA plate reader | PCR thermocycler Shaking incubator Multichannel pipette | PCR thermocycler Multichannel pipette *optional ELISA plate reader |
Capital outlay cost (for equipment above given in US$) | 96 well $25 000 48 well $19 000 | $17 800 | $50 000 | $10 000 | $7600 *optional ELISA plate reader add $5000 | $7600 | $5500 *optional ELISA plate reader add $5000 |
Hazardous chemicals | - | - | - | Ethidium bromide | TMAC, H2SO4, SDS | SDS | SDS |
Protocol run time | 1 hr 45 mins | 2 hrs | 1 hr 35 mins | ~4 hrs 30 mins | ~5 hrs 30 mins | ~16–18 hrs | ~6 hrs 30 mins |
Number of steps | 1 | 2 | 1 | 2 | 17 | 16 | 15 |
Primers/Probes required (for detection of 3 kdr alleles) | 2 PCR primers 3 fluorescently labelled probes | 2 PCR primers 3 fluorescently labelled probes | 2 PCR primers | 5 PCR primers | 2 PCR primers (one biotin labelled) 3 SSOPs (digoxigenin labelled) | 2 PCR primers 3 SSOPs (digoxigenin labelled) | 2 PCR primers 2 reporter primers (5'phosphorylation and 3'fluorescein labelled) 4 detector primers (biotin labelled) |
Number of tubes/wells/membranes required per sample | 2 | 2 | 1 | 2 | 3 | 3 | 4 |
Running cost (per sample for three alleles) | $1.72 | $1.72 | $0.62 | $0.92 | sim $1 | $1.6 | $1.74 |