How do I get started?
In MSI by PCR, DNA is first isolated from a tumor biopsy or resection sample and typically paired with a nonmalignant
sample source for comparison. Following DNA isolation, specific mononucleotide sequences within the samples are
amplified using polymerase chain reaction performed on a thermocycler. Microsatellite markers can be amplified in
multiple, parallel reactions or multiplexed in a single reaction. When multiplexed primers for individual markers are
fluorescently labeled, many different fragments of similar sizes can be detected in the same reaction.
Following amplification, the amplified fragments are resolved by size on a capillary electrophoresis instrument. Data
analysis is then performed using specialized software developed for fragment analysis.
- DNA purification system (automated or manual)
- DNA amplification reagents (or commercially available reagents)
- Marker sequence-specific DNA primers, fluorescently labeled
- Thermal stable DNA polymerase
- Salts and buffers to stabilize the polymerase and support extension from the primer
- Capillary Electrophoresis Instrument capable of multi-dye fluorescence detection and consumables
- Fragment Analysis Software