Primer Pairs and PCR Conditions for Screening of Mutation in H and Se Genes
. | Name . | Primers (5′-3′) . | Inactivated Allele Detected . | Annealing Temperature . | Fragment Sizes . | Enzyme . |
---|---|---|---|---|---|---|
H gene | mk1 | CTCGAATTCCTCAGCCTCAGAGCATTTG EcoRI | (h4) | 60°C | 1,170 bp | — (Eam1105 I) |
mk2 | CTCAAGCTTCTACTTCAGAAAGTCTCCCTG HindIII | |||||
mk3 | CTATCTGCAGGTTATGCCTCAGCTCT | h1 | 62°C | 78 bp | Xba I | |
mk4 | AACCAGTCCATGGCCTGCCGGAGG | |||||
mk5 | GCAACCACACCATTATGACCATTG | h2 | 62°C | 116 bp | Xho I | |
mk6 | GATCTTCAGGAACTCAGAGTCTCG | |||||
mk7 | AAGGGTGTGGTGGGCGACAGGGCC | h3 | 62°C | 101 bp | Apa I | |
mk8 | TCCATGCCGTTGCTGGTGACCACGA | |||||
mk9 | TTCACGACTGGATGCGGAGGTG | h5 | 62°C | 126 bp | ApaLI | |
mk10 | GCAGGGTGAACTCTCTGCGGATC | |||||
mk11 | AACTTCACCCTGCCAGACTCTGAG | h5 | 62°C | 125 bp | BstXI | |
mk12 | GGCTCTCAAGGCTTAGCCAATGTC | |||||
Se gene | tk1 | CTCGAATTCGGGCCTCCATCTCCCAGCTAAC EcoRI | 65°C | 1,170 bp | ||
tk2 | CTCAAGCTTGCTTCTCATGCCCGGGCACTC HindIII | |||||
tk5 | CAGGATCCCCTGGCAGAACTACCACATTAA | sej | 65°C | 98 bp | Ase I | |
tk6 | AGCAGGGGTAGCCGGTGAAGCGGACGTACT | |||||
tk7 | AACGACTGGATGGAGGAGGAATACCGCAGC | Se2, sej | 65°C | 80 bp | Alu I | |
tk8 | AAGGTCCAGGAGCAGGGGTAGCCGGTGAAG | |||||
Le gene | sn1 | CTCGAATTCTAAGCAGGAGATTGTCATCACTGACC EcoRI | 60°C | 1,615 bp | ||
sn2 | CTCAAGCTTCGTGCCGTGATGATCTCTCTGCAC HindIII | |||||
sn3 | CCATGGCGCCGCTGTCTGGCCGCCC | le1, le2 | 62°C | 93 bp | Msp I | |
sn4 | AGTGGCATCGTCTCGGGACACACG | |||||
sn6 | CGCTCCTTCAGCTGGGCACTGGA | le2 | 60°C | 109 bp | HindIII | |
sn7 | CGGCCTCTCAGGTGAACCAAGAAGCT | |||||
sn8 | ACTTGGAGCCACCCCCTAACTGCCA | le1 | 70°C | 206 bp | Pvu II | |
sn9 | TGAGTCCGGCTTCCAGTTGGACACC |
. | Name . | Primers (5′-3′) . | Inactivated Allele Detected . | Annealing Temperature . | Fragment Sizes . | Enzyme . |
---|---|---|---|---|---|---|
H gene | mk1 | CTCGAATTCCTCAGCCTCAGAGCATTTG EcoRI | (h4) | 60°C | 1,170 bp | — (Eam1105 I) |
mk2 | CTCAAGCTTCTACTTCAGAAAGTCTCCCTG HindIII | |||||
mk3 | CTATCTGCAGGTTATGCCTCAGCTCT | h1 | 62°C | 78 bp | Xba I | |
mk4 | AACCAGTCCATGGCCTGCCGGAGG | |||||
mk5 | GCAACCACACCATTATGACCATTG | h2 | 62°C | 116 bp | Xho I | |
mk6 | GATCTTCAGGAACTCAGAGTCTCG | |||||
mk7 | AAGGGTGTGGTGGGCGACAGGGCC | h3 | 62°C | 101 bp | Apa I | |
mk8 | TCCATGCCGTTGCTGGTGACCACGA | |||||
mk9 | TTCACGACTGGATGCGGAGGTG | h5 | 62°C | 126 bp | ApaLI | |
mk10 | GCAGGGTGAACTCTCTGCGGATC | |||||
mk11 | AACTTCACCCTGCCAGACTCTGAG | h5 | 62°C | 125 bp | BstXI | |
mk12 | GGCTCTCAAGGCTTAGCCAATGTC | |||||
Se gene | tk1 | CTCGAATTCGGGCCTCCATCTCCCAGCTAAC EcoRI | 65°C | 1,170 bp | ||
tk2 | CTCAAGCTTGCTTCTCATGCCCGGGCACTC HindIII | |||||
tk5 | CAGGATCCCCTGGCAGAACTACCACATTAA | sej | 65°C | 98 bp | Ase I | |
tk6 | AGCAGGGGTAGCCGGTGAAGCGGACGTACT | |||||
tk7 | AACGACTGGATGGAGGAGGAATACCGCAGC | Se2, sej | 65°C | 80 bp | Alu I | |
tk8 | AAGGTCCAGGAGCAGGGGTAGCCGGTGAAG | |||||
Le gene | sn1 | CTCGAATTCTAAGCAGGAGATTGTCATCACTGACC EcoRI | 60°C | 1,615 bp | ||
sn2 | CTCAAGCTTCGTGCCGTGATGATCTCTCTGCAC HindIII | |||||
sn3 | CCATGGCGCCGCTGTCTGGCCGCCC | le1, le2 | 62°C | 93 bp | Msp I | |
sn4 | AGTGGCATCGTCTCGGGACACACG | |||||
sn6 | CGCTCCTTCAGCTGGGCACTGGA | le2 | 60°C | 109 bp | HindIII | |
sn7 | CGGCCTCTCAGGTGAACCAAGAAGCT | |||||
sn8 | ACTTGGAGCCACCCCCTAACTGCCA | le1 | 70°C | 206 bp | Pvu II | |
sn9 | TGAGTCCGGCTTCCAGTTGGACACC |