Microsatellites are some very short repeats of one to four bases pairs long in DNA. The number of tandem repeats are highly polymorphism, depend on the species, on the individuals and even on the alleles. Because microsatellites polymorphism are very common and often appear in all the genome, they are ideal to be use in genetic mapping.
For the microsatellite polymorphism genotyping, we first running a PCR with a fluorescent primer to amplify short sequence containing the microsatellite. Then, by capillary electrophoresis, we can know the size of the fragments.
We use four different fluorochromes:
- 6-FAM
- VIC
- PET
- NED
- LIZ (ladder of 500 bp)
The same fluorochrome can be used more than once when there is a difference in length of more than 100 nucleotides between the fragments. Given that, we can multiplex up to 12 different markers per person using the same sample.
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