AGAROSE-BASED SYSTEM FOR SEPARATION OF SHORT TANDEM REPEAT LOCI

Short tandem repeats (STRs) are traditionally analyzed on large polyac rylamide electrophoresis gels. We demonstrate in this study that a sma ll (10-cm-long, 1-mm-thick) agarose gel is sufficient for analysis of multiplexed samples for several commonly used STR loci. A system was d eveloped using a high-resolution agarose, MetaPhor(R). Within an hour of electrophoresis, sufficient resolution was obtained to allow discri mination of triple-multiplexed STR loci. We show that this agarose is capable of resolving di- as well as tetranucleotide ladders. Using PCR conditions similar to those routinely used with sensitive detection s ystems, we found that direct staining of gels with SYBR(R) Green I sta in was comparable with sillier staining, autoradiography of fluorescen tly tagged primers for sample detection and was considerably easier Th is procedure significantly simplifies STR analysis and is much faster than many standard protocols.