PCR Primers

   
 

RT-PCR (mouse genes):

 

 

   
Beta-actin: (115bp) 5’-TCCGTAAAGAATCTCATGCC
  5’-ATCTTCATCCTCCTAGGAGC
   
Pax1: (273bp) 5’-CACATTCAGTCAGCAACATCCTG
  5’-TGTATACTCCCTGCTGGTTGGAA
   
Pax3: (454bp) 5’-AGGAGGCGGATCTAGAAAGGAG
  5’-TGTGGAATAGACGTGGGCTGGTA
   
Sim1: (219bp) 5’-GGAGCCCGAGACACGATGAAAG
  5’-AAGGGGCTGGTTCGACTGGTGT
   
Pax7: (395bp) 5’-GACAAAGGGAACCGTCTGGATGA
  5’-TGTACTGTGCTGCCTCCATCTTG
   
Mtwist: (312bp) 5’-GCGCAAGATCATCCCCACGCTGC
  5’-CCAGCTCCAGAGTCTCTAGACTGT
   
Pax9: (450 bp) 5’-GATGGCGTGTGCGACAAGTAC
  5’-CTGGCCGTACTTGGCTTCCTG
   
Myf5: (190bp) 5’-TACATCAGGACAGTAGATGCTGTC
  5’-CATCCGCTACATTGAGAGCCTC
   
MyoD: (371bp) 5’-CGCTCCAACTGCTCTGATGGCA
  5’-TGCTGCTGCAGTCGATCTCTCA
   
Bmp2: (436bp) 5'-CGGAGACTCTCTCAATGGAC
  5'-GTTCCTCCACGGCTTCTAGT
   
Bmp4: (468bp) 5'-CTCCCAAGAATCATGGACTG
  5'-AAAGCAGAGCTCTCACTGGT
   
Sfrp1: (360bp) 5’-CGAGATGCTCAAATGTGACAAGT
  5’-CCCATGATGAGAAGTTGTGGCT
   
Sfrp2: (465 bp) 5’-AGACATGCTGGAGTGCGACCGTT
  5’-CACTGGGAAACTAGCATTGCAGC
   
Sfrp3: (270 bp) 5’-ATTCTCATCAAGTACCGCCAC
  5’-TGCCTTTAGAATTTCCTTCAC
   
Sfrp4: (453 bp) 5’-TCATGAAGATGTATAACCACAG
  5’-GCCACTCATAACACATGATTAG
   
Ptc1: (515bp) 5’-AAGGCAGATAATCTCGAGACC
  5’-GCCAACTTCGGCTTTATTCAG
   
Hip: (294bp) 5’-GCAGAATTGCCAAGTGTGAGC
  5’-ATAGACTCGAACTGTCCCAG
   
Gli1: 5’-ATCACCTGTTGGGGATGCTGGAT
  5’-GGCGTGAATAGGACTTCCGACAG
   
Gli2: 5’-GTTCCAAGGCCTACTCTCGCCTG
  5’-CTTGAGCAGTGGAGCACGGACAT
   
Gli3: 5’-AGCAACCAGGAGCCTGAAGTCAT
  5’-GTCTTGAGTAGGCTTTTGTGCAA
   
Sox9: (1081bp) 5’-AAGATAAGTTCCCCGTGTGC
Semba et al., 2000 5’-GTAGTGAGGAAGGTTGAAGG
   
CyclinD1: 5’-ATGGTAGCTGCTGGGAGCGTGGT
  5’-CTTTGCGGGTGCCACTACTTGGT
   
CyclinD2: 5’-GCAAGCATGCGCAGACCTTCATC
  5’-ATGACGAACACGCCTCTCTCTTG
   
CyclinE1: 5’-CCACAGCAACATGAAAGAAGAAG
  5’-TGCAGTCCTAGGGTCTTCAAGCT
   
CyclinE2: 5’-AAGCAACATGGCCAGCCCAACCA
  5’-TGTGAACATGCCCAGCTTAAATC
   
Brachyury: (500bp) 5’-TGCTGCCTGTGAGTCATA
Arnold et al., 2000 5’-ACAAGAGGCTGTAGAACATG
   
Tcf1: (332bp) 5’-GAAGGAGGCTAAGAAGCCAGT
  5’-GCCTTCTCCGGGTAAGTACCG
   
Lef1: (514bp) 5’-CACAACTGGCATCCCTCATCC
  5’-CCATCTCCAGAAGAGGGCCTG
   
Tcf3: (438bp) 5’-CTGCCATCGTCTCCCCCATTG
  5’-GCTTCTTGCTCTTAGAGGCCAG
   
Tcf4: (424bp) 5’-CAGCTCAAAGCATCAGGACTC
  5’-CTGTTGATCAAGGCCAAAGCGC
   
BTEB2: (381bp) 5’-GGTGTATTCAACAGCTGGTAC
  5’-ATAGACGAGACAGTGCCTCAG
   
Cdx1: (370bp) 5’- GTAAGACCCGAACCAAGGAC
Prinos et al., 2001 5’-CAGGATATCCTAGGGTAGAAACTCCTCCTTGACG
   
Cox2: (306 bp) 5’-GTGATCGAAGACTACGTGCA
  5’-GCTTGTACAGCAATTGGCAC
   
Eda: (139bp) 5’-ATTGTCTTATGCTAGGTTCATGAGACTT
  5’-CCCATCTCCTCAAAAAGAAATACATT
   
Wnt6: (468bp) 5’-CATGGATCCTACCAGCATCTGCAG
  5’-GCTGCGCATCCATAAAGAGTCTTG
   
Wnt11: (385bp) 5’-GAGCTCATGCGCACCATCGTGCAC
  5’-GTGTAGACGCATCAGTTTATTGGC
   
Troy: (460bp) 5’-CAAGGTCCTACCTCTACACA
Kojima et al., 2000 5’-AAGGTTCACCTTGCTGGTAC
   
Selp: (309 bp) 5’-GTGACTGCAGCCTTTGGTTA
  5’-TGCAGTGCAACTCAAGCTAG
   
Arl4: (199 bp) 5’-GGCAGTACCTTTTATATTGG
  5’-GGAGAAGAGACCACTTTAAG
   
Sim2: (260bp) 5’-AGCCGAGGCGCGATGAAGGAGAA
  5’-ATGGGATCCCAGCTCTTTTG
   
Mc3R: (~700bp) 5’-GGCAACCTGCACTCTC
  5’-CATGCCCAAGTTCATGC
   
Tbr1: (~700BP) 5’-GACAACCTGGAGAGAAG
  5’-AACTGGTTTTGTGCC
   
G3PDH (515 bp) 5’-ACCACAGTCCATGCCATCAC
  5’-TCCACCACCCTGTTGCTGTA
   

 

PCR primers used to generate in situ probes:

   
Pax1 3’UTR probe (660 bp): 5’- CGAATTCCTCACACACACTTTTCTCAC
  5’-GCGGATCCACCAGTGCGCCTTAACTT.
   
Pax3 3’UTR probe (547bp): 5’-CGAATTCTGGAAGGGAGAGAGATCAC
  5’-GCGGATCCTTCATGTCTAGTCTGTGGA
   
Pax9 probe (450 bp): 5’-GATGGCGTGTGCGACAAGTAC
  5’-CTGGCCGTACTTGGCTTCCTG
   
Arnt2 probe: 5’-CAGAACCCCTACTCTGATGA
  5’-GATAATAGCGTCTGGGCAGT
   
Trh probe: 5’-GTGCCAACCAAGACAAGGAT
  5’-CAATCCTACCTTTCTGAGG
   
Brn2 probe: 5’-AAGCAATTCAAGCAGAGGCG
  5’-CTACTTCATTGCCTGGGTAC
   
Sox9 probe (1081bp): 5’-AAGATAAGTTCCCCGTGTGC
Semba et al., 2000 5’-GTAGTGAGGAAGGTTGAAGG
   

 

PCR primers for genotyping mutant animals:

   
Sim1 genotyping:  
Sim1 wt allele: (287bp) 5’-ATACCCAGGGCTGTGACGAG
  5’-AATGCTGGAAGGACTCCCGA
   
Neo allele: (189bp) 5’-CTCGGCAGGAGCAAGGTGAGATG
  5’-GTCAAGACCGACCTGTCCGGTGC
   
Sim2 genotyping:  
Sim2 wt allele: (135bp) 5’-AGCCGAGGCGCGATGAAGGAGAA
  5’-TGGACGCTTTGTCCAGCTGCGAG
   
Neo allele: (189bp) 5’-CTCGGCAGGAGCAAGGTGAGATG
  5’-GTCAAGACCGACCTGTCCGGTGC
   
Sim1-tau-LacZ genotyping: (also for Sim1-tau-LacZ X Sim1)
Sim1 wt allele: 5’-CCGAGTGTGATCTCTAATTGA
  5’-TAGGCACAGACGCTTACCTT
   
tau-LacZ allele: 5’-AGTGACCAGCGAATACCTGTTCC
  5’-ACTACGCGTACTGTGAGCCAGAG
   
Gli2 genotyping: (Mo et al. 1998)
Gli2 wt allele: 5’-AAACAAAGCTCCTGTACACG
  5’-CACCCCAAAGACTGTGTTTT
   
Gli2 mutant allele: 5’-ATGCCTGCTCTTTACTGAAG
   
Gli3 XtJ genotyping: (Maynard et al 2002)
Gli3 wt allele: (193bp) 5’-GGCCCAAACATCTACCAACACATAG
  5’-GTTGGCTGCTGCATGAAGACTGAC
   
Gli3 mutant allele: (580bp) 5’-TACCCCAGCAGGAGACTCAGATTAG
  5’-AAACCCGTGGCTCAGGACAAG
   
Shh genotyping: (Chiang et al., 1996)
Gas1 genotyping:  
Gas1 wt allele: (350bp) 5’-ATCTCGGCGCTTATCCAGCTCAAC
  5’-CATCGCACACGCAGTCGTTGAGCA
   
Neo allele: (189bp) 5’-CTCGGCAGGAGCAAGGTGAGATG
  5’-GTCAAGACCGACCTGTCCGGTGC
   
Single-LacZ allele (for transgenic reporters):
LacZ: (500-600bp) 5’-ACCAGAAGCGGTGCCGAAAA
  5’-CCCGTAGGTAGTCACGCAAC