Analysis of Horse Myostatin Gene and Identification of Single Nucleotide Polymorphisms in Breeds of Different Morphological Types
Table 2
Primers, PCR conditions, amplified regions, and use of the PCR products.
Primer pair
F: Forward sequence (-) R: Reverse sequence (-)
Primer annealing regions
Amplified region (bp)*
PCR
Use of the PCR products
1F: TCAGGGAAACAAGTTTCTCAAAT
Promoter
1–484
58/1.5/45/
Sequencing,
1R: TGCTCCACAATGAATCTCG
Promoter
(484)
E
PCR-RFLP
2F: TGAATCAGCTCACCCTTGAC
Promoter
369–727
62/0.8/45/
Sequencing
2R: CCAGCAACAATCAGCATAAA
Exon 1
(360)
E
3F TGTGCTGATTCTTGCTGGTC
Exon 1
710–947
58/1.5/45/
Sequencing
3R: ATCAATCAGTTCCCGGAGTG
Exon 1
(238)
E
4F: GACCCGTCAAGACTCCTACA
Exon 2
2982–3226
58/1.0/45/
Sequencing
4F: TGGGAAGGTTACAGCAAGA
Exon 2
(245)
E
5F: AGGCCAATTACTGCTCTGGA
Exon 3
5430–5744
58/1.5/45/
Sequencing
5R: ATACTCTAGGCTTATAGCCT
3’ UTR
(315)
E
6F: CACTCCGGGAACTGATTGAT
Exon 1
928–3098
58/0.8/130
Sequencing
6R: CGCCTGGGTTCATGTCAAGT
Exon 2
(2171)
/T
7F: AGGCAGGCACATTGCTTAAT
Intron 1
1808–2287
58/0.8/45/
Sequencing
7R: GAATGTTATATTCAGGCTATCTCAA
Intron 1
(480)
E
8F: AAATGTGACATAAGCAAATGATTAG
Intron 2
5152–5534
62/0.8/45/
Sequencing
8R: AGCAGGGGCCTGCTGAACCTCTGGG
Exon 3
(383)
E
9F: TGCAAAATTGGCTCAAACAG
Exon 2
3135–5361
55/0.8/130
Sequencing
9R: CAGCATCGAGATTCTGTGGA
Exon 3
(2227)
/T
10F: CCCCCAGAAGAGTGTCAAAT
Intron 2
3627–4822
60/0.8/130
Sequencing
10R: TCTTTACTTGGGGAAACTTGGA
Intron 2
(1196)
/T
11
1F: TCAGGGAAACAAGTTTCTCAAAT
Promoter
1–204
53/1.2/35/
PCR-RFLP
11R: ACTTCCTCAGAAATTAAGATTTAAT
Promoter
(204)
E
accoding to GenBank accession number GQ183900. §Annealing temperature (), [MgCl2], extension time (s), Taq DNA polymerase (E = EuroTaq; T = TaKaRa). pairs 1 and 2 were designed based on conserved sequences of the published cattle, pig, and humans MSTN promoter regions (GenBank accession numbers AJ310751, AJ133580, and AX058992, respectively). pairs 3–10 were designed based on horse MSTN mRNA sequence (GenBank accession number AB033541).
