title: soil eukaryotic 18S SSU rRNA diversity in three neotropical broadleaf forests
design: PCR primers are V4F = CCAGCASCYGCGGTAATTCC and V4R = ACTTTCGTTCTTGATYRA, also known as TAReuk454FWD1 (forward) 5'-CCAGCA(G/C)C(C/T)GCGGTAATTCC-3' and TAReukRev3 (reverse) 5'-ACTTTCGTTCTTGAT(C/T)(A/-G)A-3'.  PCR mix is (volumes in microliters) 19.75 H20, 2.5 10x Buffer, 0.5 dNTP, 0.5 V4F, 0.5 V4R, 0.25 polymerase, 1 template. Cycling conditions (with Hot Start Taq from Qiagen) are 1) 95C 5 min, 2) 94C 30 sec, 3) 47C 45 sec, 4) 72C 60 sec, repeat 2-4 = 29x, 5) 72C 5 min.