using reverse transcriptase superscript u2122 iii one step rt pcr system with platinum u00ae taq dna polymerase kit from invitrogen pcr amplification was performed in a 100 u03bcl reaction volume using 2 u03bcl cdna 1 u03bcl 10 mm dntps 10 u03bcl taq polymerase buffer 2 5 u taq polymerase from promega madison wis and 20 pmol 5 u2032 or 3 u2032 primers in h 2 o specific primers used herein were heavy chain forward a mixture of 5 u2032 ctt ccg gaa ttc sar gtn mag ctg sag sag tc 3 u2032 seq id no 10 5 u2032 ctt ccg gaa ttc sar gtn mag ctg sag sag tcw gg 3 u2032 seq id no 11 5 u2032 cct ccg gaa ttc cag gtt act ctg aaa gwg tst g 3 u2032 seq id no 12 5 u2032 ctt ccg gaa ttc gag gtc car ctg caa car tc 3 u2032 seq id no 13 5 u2032 ctt ccg gaa ttc cag gtc caa ctv cag car cc 3 u2032 seq id no 14 5 u2032 ctt ccg gaa ttc gag gtg aas stg gtg gaa tc 3 u2032 seq id no 15 5 u2032 ctt ccg gaa ttc gat gtg aac ttg gaa gtg tc 3 u2032 seq id no 16 heavy chain reverse 5 u2032 gga aga tct gac att tgg gaa gga ctg act ctc 3 seq id no 17 light chain forward 5 u2032 gg gag ctc gay att gtg mts acm car wct mca 3 u2032 seq id no 18 and light chain reverse 5 u2032 ggt gca tgc gga tac agt tgg tgc agc atc 3 u2032 seq id no 19 wang et al universal pcr amplification of mouse immunoglobulin gene variable regions the design of degenerate primers and an assessment of the effect.