Volume 47, Issue 3 (May 2002)
Allele Frequencies for Three STR Loci RT24, RT09, and BM1225 in Northern New England White-tailed Deer
Muscle samples (~1 cm3) were obtained from harvested animals in four distinct locations in each state (total sample size = 362). Maine samples were collected during the 1998 deer hunting season (November 1–31, 1998), New Hampshire samples were collected during the 1999 hunt (November 13–19, 1999), and Vermont samples were collected during the 1999 hunt (November 13–19, 1999). DNA was extracted using the QIAmp™ tissue kit (Quiagen, Valencia, CA) as per the manufacturer's instructions (1). One to two µL of extracted DNA was used in a 25 µL multiplex PCR reaction containing 10 mM KCl, 20 mM Tris pH 9.5, 10 mM (NH4)2SO4, 1 mM MgCl, 4 mM dNTP's (Promega, Madison, WI), 0.75 units of Taq DNA Polymerase (GibcoBRL, Grand Island, NY), and 0.3 µM of each primer (Operon, Almeda, CA). Three STR loci were examined: BM1225 [(2), primers: BM1225f ttt ctc aac aga ggt gtc cac BM1225r acc cct atc acc atg ctc tg], RT09 [(3), primers: RT9f tga agt tta att tcc act ct, RT9r cag tca ctt tca tcc cac at] and RT24 [(3), primers: RT24f tgt atc cat ctg gaa gat ttc ag, RT24r cag ttt aac cag tcc tct gtg]. PCR was performed in a HYBAid OMN-E Personal Thermal Cycler™ (HYBAid, Middlesex, UK) for 30 cycles (94°C, 30s; 54°C, 45 s; 70°C, 2 min) and a final extension (72°C, 10 min). PCR products were electrophoresed and analyzed with an ABI 377™ automated DNA sequencer as per the manufacturer's instructions (Applied Biosystems International, Foster City, CA). Electropherograms were interpreted using Genescan™ (ver. 3.1) and Genotyper™ (ver. 2.1) software (Applied Biosystems International, Foster City, CA). Allele frequencies and heterozygosities were calculated with Genepop (ver. 3.1c) (4) and Hardy Weinberg tests were performed with GDA (ver. 1.0d16c) (5).