How are DNA microsatellites amplified by polymerase chain reaction (PCR)?

Methods: DNA microsatellites were amplified by polymerase chain reaction (PCR) using a pair of specific primers tagged with fluorescence to yield products of 160–200 base pairs in length.

How to detect length polymorphism of a microsatellite marker?

The length polymorphism of a microsatellite marker is commonly detected through polymerase chain reaction (PCR) amplification using pairs of specific primers flanking tandem arrays of microsatellite repetitions and proceeded by electrophoresis ( 11 ). Horizontal electrophoretic analyses are commonly performed using agarose gels.

Does the banding sequence of a microsatellite (Ca)n exhibit allelic dimorphism?

Although the banding patterns were observed and measured using gel electrophoresis, which suggests that the banding sequence of this microsatellite (CA)n exhibits allelic dimorphism, it is still not the best technique to determine the amplicon size; therefore, the size of the CA-repeats.

Can capillary electrophoresis determine the length of polymorphisms of microsatellite markers?

In our analysis, capillary electrophoresis has also been used to determine the length of polymorphisms of microsatellite markers ( 17 ). This method requires highly sophisticated instruments and fluorescently tagged primers, which are quite expensive. The results are confirmed to be precise and reliable.

How is a PCR product electrophoresis optimisation performed?

First, the PCR products were run in 2%, 3%, and 4% (weight/volume) LMA gel in 1X Tris/Borate/EDTA (TBE) buffer at 100V at 90 minutes for gel electrophoresis optimisation. Then the PCR products were electrophoresed on a 4% LMA gel (Fisher Scientific) using 1X TBE buffer at 100 V for 1 hour and 25 minutes.

How are microsatellite loci amplified by PCR?

Microsatellite loci are amplified by PCR using fluorescently labeled forward primers and unlabeled reverse primers. A pair of microsatellite primers was custom synthesised at First BASE Laboratories Sdn Bhd (Selangor, Malaysia) and utilised in the study for amplification of PCOS markers.