The MLPA procedure includes the separation of MLPA fragments on a capillary electrophoresis device. The table below lists some of the more common capillary electrophoresis devices supported by Coffalyser.Net.
| Supplier | Device(s) |
|---|---|
| Applied Biosystems | SeqStudio Genetic Analyzer |
| SeqStudio Flex Series Genetic Analyzer | |
| 3500 Series Genetic Analyzer | |
| 3730 Series Genetic Analyzer | |
| 3130 Series Genetic Analyzer | |
| Hitachi | Compact CE Sequencer DS3000 |
| Promega | Spectrum Compact CE System |
| SCIEX/Beckman Coulter | GenomeLab GeXP |
| CEQ 8800 Genetic Analysis System | |
| CEQ 8000 Genetic Analysis System |
A good capillary electrophoresis device must:
- Be supported by Coffalyser.Net; see the Reference Manual for a full list of supported devices.
- Use denaturing conditions.
- Be compatible with 6-FAM or Cy5.0 labels used for our primers, and have a second channel suitable for a size marker.
- Be capable of separating fragments in the 50–500 nt range with sufficiently high resolution to separate MLPA fragments.
- Have a linear relationship between peak height and fragment concentration (within a certain signal range).
- Provide clean and reproducible peak patterns (e.g. no excess noise, extra peaks or inconsistent signal heights).
Unsuitable capillary electrophoresis devices
Capillary electrophoresis devices that use non-denaturing conditions are not suitable. Secondary structures will alter the relative mobility of MLPA reaction products, which can lead to different apparent sizes or even to position swaps of fragments in the electropherogram. Devices that this applies to, which are not suitable for MLPA, include the Agilent (formerly Advanced Analytical/AATI) Fragment Analyzer, the Agilent 2100 Bioanalyzer, the Agilent TapeStation systems, the Caliper LabChip GX/GXII and the QIAGEN QIAxcel.