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Table 1 Components and conditions used in PCR-RFLP of rs3917655, rs3917657, rs3917739, rs3917843 and rs223530

From: Putative functional non-coding polymorphisms in SELP significantly modulate sP-selectin levels, arterial stiffness and type 2 diabetes mellitus susceptibility

SNP

Primer sequence

PCR conditions

Amplicone Size (bp)

RFLP

Restriction enzymes

Incubation

conditions

Product after digestion (bp)

Ancestral

Variant

rs3917655

5’TGTCCACTTTGACCCTCCCA3’

5’AGGGCAGAAAAGGAAACTATGTG3’

Initial denaturation at 95 °C (7 min)

30 s at 95 °C

30 s at 58 °C

30 s at 72 °C

Final elongation at 72 °C for 7 min

405

PvuII

At 37 °C for 2 h

249

405

156

rs3917657

5’ATCTTCTGGGACTGATCTGGA3’

5’CCTGCCTGGTTCCTCCATAG3’

Initial denaturation at 95 °C (7 min)

30 s at 95 °C

30 s at 60 °C

30 s at 72 °C

Final elongation at 72 °C for 7 min

516

TfiI

At 65 °C for 2 h

265

265

251

199

52

rs3917739

5’AAAGCCCAGAGCAAAGAGGTAGT3’

5’CCCTCCCTTCCCACCTTAACT3’

Initial denaturation at 95 °C (7 min)

30 s at 95 °C

30 s at 60 °C

30 s at 72 °C

Final elongation at 72 °C for 7 min

546

TfiI

At 65 °C for 2 h

546

328

218

rs3917843

5’ATTACATGCAATGCCTGCCT3’

5’GGGGCATACTGTCCCTTTTTGA3’

Initial denaturation at 95 °C (7 min)

30 s at 95 °C

30 s at 59 °C

30 s at 72 °C

Final elongation at 72 °C for 7 min

578

BsaWI

At 60 °C for 2 h

329

578

249

rs2235302

5’GCCAACCTGTGAGGGTAGGAT3’

5’ACCACTGTCCGCCTTATAAACT3’

Initial denaturation at 95 °C (7 min)

30 s at 95 °C

30 s at 57 °C

30 s at 72 °C

Final elongation at 72 °C for 7 min

511

EciI

At 37 °C for 2 h

441

511

70

  1. PCR and digestion products were analyzed on 1.5 and 2.5% agarose gel pertained with EtBr, respectively