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Table 3 Primer sequences used for mutation analysis of individual exons/introns of the canine SAG gene

From: Screening of the arrestin gene in dogs afflicted with generalized progressive retinal atrophy

Primer

Location

Forward primer, reverse primer (5'–3')a

PCR conditions [T-°C/MgCl2-mM]

PCR amplicon length (bp)

Restriction enzymes for SSCP analyses

UTR1F

exon 1

GGGCAACCCTGTCCAGGT

54/1.0

699

Nla III/Rsa I

UTRI-1aR

intron 1

TCTATCATGACGGGACGCCT

   

UTRI-1aF

intron 1

GAAAATGATATTTGCAAAGCAG

50/1.0b

283

Tru 1I

UTRI-1aR

intron 1

TCTATCATGACGGGACGCCT

   

1-IF

intron 1

CTAATGGGCACACAGCATCTC

53/1.0

249

Pvu III; Nla IIIc

2-I2R

intron 2

TTCTGTTAAGCCACTCACTTC

   

3-IF

intron 2

TGTTTTTATCTAACACTGACTACTTC

48/1.0b

224

Nla III

3-IR

intron 3

AAATAACAAAGTAGCAGCTGTC

   

4-IF

intron 3

AACTGCAGATAAATATATGAAG

52/1.0

189

Alu I

4-IR

intron 4

AAAGTTCTTTCCTAGCACTAAG

   

5-IF

intron 4

GGTTACCCCATGTTCACTTG

56/1.0

343

Mnl I

5-IR

intron 5

GCTCCTGGTCACACTGCAAG

   

6-IF

intron 5

CAAGTTTTACACACTGAGTGC

55/1.0

213

Alu I

6-IR

intron 6

ATTTTCCCCAGAGAAAAGGCTA

   

7-IF

intron 6

CGGGAAGGGAGGTGCTGA

58/1.0

233

Rsa Ic

7-IR

intron 7

TCGCAGGCCACAGAGGAGAAG

   

8-IF

intron 7

ATCACAGCGTGAGTACGGGGAG

55/1.0

288

RsaI; Pst Ic

8-IR

intron 8

CAGCACCCACAGCAGATTG

   

9-IF

intron 8

CCTAGAAAGCCATGAGATTAA

55/1.5

214

Tru I1; Sty Ic

9-IR

intron 9

GACCAGACTGAGAAATTCTAG

   

10-IF

intron 9

GGCACCATGCACATGCGTG

58/1.0

235

Alu I

10-IR

intron 10

AAGCCAAGCATCCTACTTCC

   

11-IF

intron 10

GGACTGATGGTGGCTTTATG

58/1.0

266

BsuR I

11-IR

intron 11

AGCAGACCAGCACCTCCTC

   

12-IF

intron 11

TGAGGGATGTGTTCATCTAG

55/1.0

183

-

12-IR

intron 12

GCCCATGGTGTTGGCTCTTG

   

13-IF

intron 12

CATGCTTGGGACATGTCCAC

64/1.0

209

Mva I

13-IR

intron 13

TTCATAACACCTCTGAGCTAC

   

14-IF

intron 13

CTCTGCAGCCACAGCCCTTC

50/2.0

229

Ava II

14-IR

intron 14

GTGCTCAGGGACCACTAG

   

15-IF

intron 14

CCTGCTCACGATTCTCTTTC

58/1.0

244

Hph I

15-IR

intron 15

AAGCCGAAGACTGGGAGC

   

16-IF

intron 15

GATCGGTCCCTTGTTGCA

52/2.0b

347

Alu I

16-ER

Exon 16

CACAGCTGAACAGACAAACTT

   
  1. aSee EMBL accession numbers AJ426068-AJ426078 bwith 5% formamide cwith RLFP analysis