Table 1.

PCR Conditions for Amplifying 68 Equine Type I Loci with Universal Primers

Hsa UCD Symbol Locus name Anl T. °C MgCl (mm) R.E. Size (bp) Accession no.[ii]
1p36.22 NPPA natriuretic peptide precursor A502 HinfI305 AF134229
1p13.15 NGFB nerve growth factor, β532.5 RsaI158 AF134228
1q215 GBA glucosidase, β acid532 RsaI345 AF134220
1q23–q25.15 AT3 antithrombin III552187 AF134055
1q315 LAMC1 laminin, C-1502∼1400 AF134225, 6
2p2115 SPTBN1[iii] spectrin, β, nonerythrocytic 1502975 AF130782
2q12–q1415 PAX8 paired box homeotic gene 8552 HaeIII147 AF130778
2q1415 IL1b[iv] [v] interleukin-1 β551 HindIII∼906 AF130767, 8
2q24–q3218 CHRNA cholinergic receptor, nicotinic, α581405 AF130750
2q31–q3218 NEB nebulin561 PvuII∼2000 AF130774, 5
2q33–q34D CHRNG cholinergic receptor, nicotinic, γ601264 AF130751
2q34D FN1 fibronectin 15521009 AF130764
3p24.3–p24.216 RARB retonic acid receptor, β553 CfoI208AF13424
3p21.3316 GLB1 [iii] galactosidase, β-1502 RsaI429 AF130765
3q21–q2416 CP ceruloplasmin552.5∼1500 AF134057
3q21–q2416 RHO [iii] rhodopsin552 RsaI327 AF130780
3q2819 SST[vi] somatostatin501931 AF130783
4q223 ADH2 alcohol dehydrogenase 2521295 AF133272
4q223 ADH3 alcohol dehydrogenase 3522>2000 AF134056
4q282 FGG fibrinogen, γ polypeptide532∼1100 AF134218, 9
5q32–q3414 ADRB2 adrenergic receptor, β-2552 HinfI354 AF130746
6p24–p2320 EDNI[iii] endothelin-1582266 AF130760
6q1320 COL9a1[iii] collagen, type IX, α-1502292 AF130755
6q21.1–q2310 CGA chorionic gonadotropin, α chain454242 AF130749
6q21–q22.310 COL10[iii] collagen, type X, α 1502 PVuII365 AF130754
6q25.314 SOD2ps[viii] superoxide dismutase 2552154 AF130781
7p15–p144 TCRG t cell antigen receptor, γ subunit422.5 HinfI252 AF134235
7q11.213 ELN[iii] elastin581263 AF130761
7q21.1113 GUSB glucuronidase β532 Sau3AI382 AF134223
7q364 EN2 Engrailed 2532 PvuII189 AF134060
8q24.2–q24.39 TG thyroglobulin581749 AF130785
9p2223 IFNA1 interferon, α-1532 PvuII373 AF135017
9p2123 IFNB1 interferon, β-1552 HaeIII450 AF134227
9p1323 CNTFR ciliary neurotrophic f. recept.572.5456 AF134058
9q3125 TXN thioredoxin552 HinfI∼780 AF134237
9q3425 GRP78 glucose-regulated protein552 PvuII∼740 AF134222
10p1329 VIM vimentin572∼1230 AF135018
11p15.57 HBB hemoglobin, β502.5171 AF134224
11p15.3–p15.17 PTH parathyroid hormone552311 AF134233
11cen–q1312 ADRBK1 adrenergic receptor β kinase I531328 AF134059
11q1312 CD20[iii] CD20 antigen5521007 AF130748
11q237 DRD2 dopamine receptor D2502.5∼1300 AF134061, 2
11q23.37 THY1 Thy-1 T-cell surface antigen502∼800 AF134236
12p12.19 KRAS2ps[viii] Kirsten murine sarc. 2 viral onc., ps.581329 AF130769
12q13.3D PFKM phosphofructokinase m602∼1600 AF134230, 1
12q14D IFNG Interferon γ452325 AF130766
12q24.26 TCF1 transcription factor 1581602 AF130784
13q14.1q–14.217 RB1[vii] retinoblastoma552705 AF130779
14q11.21 CHY[iii] chymase- mast cell582697 AF130752
14q121 MYH6[v] myosin, heavy pol. 6 &/or 7, see text582 RsaI∼650 AF130771, 2
15q22–qter1 CYP1A2 cytochrome P450, subf. I, polyp. 2502.5 HinfI278 AF134063
15q26.11 FES[iii] feline sarcoma virus homologue582 510 AF130763
17q11.211 EVI2A ecotropic viral int. site 2A554 156 AF130762
17q11.211 NF1 neurofibromatosis I582 380 AF130776
17q2511 P4HB prolyl-4 hydroxylase β622 460 AF130777
1711 MYL4 myosin light chain 4582 682 AF130773
18p11.326 TS[iii] thymidylate synthetase551 539 AF130786
19q1310 CKM[iii] creatine kinase, muscle581 819 AF130753
20pter–p1222 PRNP prion protein551.5 191 AF134232
20q13.222 GNAS1 guanine nuc. binding protein501.5 RsaI324 AF134221
20q13.1122 ADA adenosine deaminase501.5 ∼1350 AF135790
21q22.326 ETS2 avian erythroblastosis virus onc. 2552 HinfI∼1000 AF134064
22q13.1C CYP2D[v] cytochrome P450, subfamily IID582 980 AF130756- 9
22q13.31–qterC ARSA arylsulfatase A582 247 AF130747
Xp11.21X ALAS2 Δ-aminolevulinate synthase532 HaeIII∼800 AF133200
Xq27.1–q27.2X F9 coagulation factor IX552 453 AF133201
Xq28X BGN biglycan572 HinfI∼710 AF135019, 0
Xq28X G6PD glucose-6-phosphate dehydro.552 HinfI∼750 AF133202

[i] All primers used in this study were published by Lyons and coworkers (1997), with the following exceptions:

[ii] Multiple accession numbers indicated sequences obtained from the 5′ and 3′ ends of partially sequenced PCR products, or multiple highly homologous sequences most likely amplified from different syntenic isoforms of paralogous genes (see text).

[iii] Primers published by Venta and associates (1996).

[iv] The primers used to amplify equine IL1B were originally designed to amplify IL1A (see text).

[v] These primers amplify multiple equine fragments of different but highly homologous sequences (see text).

[vi] Amplifies a fragment of unknown sequence in addition to the equine SST gene (see text).

[vii] The forward primer used to amplify equine RB1 (5′=TTTGATATCGAAGGGTCTGAC-3′) was designed in this study and the reverse primer was as in Venta et al. (1996).

[viii] These primers amplify equine sequences lacking expected introns and therefore are thought to be horse pseudogenes from the respective loci.