From stdabb14 from shsu.edu Mon Jul 16 13:56:30 2007 From: stdabb14 from shsu.edu (Anna B. Buchman) Date: Mon Jul 16 17:09:28 2007 Subject: [Genetic-linkage] Primer Design for Microsatellites Message-ID: <3385.158.135.197.110.1184612190.squirrel@sammail.shsu.edu> Hi, I'm in the process of developing microsatellite primers to study box turtle hybridization, and I am now at the initial stage of actual primer design. Any recommendations as to what software is best for designing primers for microsats? I'm currently exploring Invitrogen's Vector NTI and B. Fairthcloth's Msatcommander (works off of primer 3.) Also, any advice on what is the best way to process and analyze samples once the microsat regions are ID'd and primers are designed? I'm going to need to do a few hundred samples, and our facilities do not have a functional sequencer. What companies out there do cheap AFLP analysis? Thanks. From stdabb14 from shsu.edu Wed Jul 25 15:00:21 2007 From: stdabb14 from shsu.edu (Anna B. Buchman) Date: Wed Jul 25 18:53:29 2007 Subject: [Genetic-linkage] Primer Design for Microsatellites Message-ID: <2824.158.135.188.13.1185393621.squirrel@sammail.shsu.edu> Hi, Thanks for all the replies to the previous posting, guys. I have now ordered the primers, and have a commercial sequencing question. I plan to fluorescently tag the primers when I verify that they work, and am looking to send the resulting tagged PCR products to a company that can do an AFLP/microsatellite variability assay on them, since we don't have an automated sequencer and I would like to avoid manually running and analyzing sequencing gels. What companies can provide that service for a reasonable price? Also, if I do end up doing it myself, any ideas on how to simplify the procedure overall? Anything radioactive is out of the question, but I have read others doing silver staining with polyacrilamide gels rather than flourescent tagging, and even running agarose gels. Thanks.