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Table 1 Sequences of PCR primers used and their PCR conditions.

From: Screening for known mutations in EIF2Bgenes in a large panel of patients with premature ovarian failure

Nucleotide change tested (gene)

Primers sequences (5'-3')

PCR conditions

C512T (EIF2B2), C547T (EIF2B2)

F: GCAAAACCGTTCTTAC

R: CCTACCCATCTCTCGTTTAT

PCR preparation: 1.5 mM MgCl2, 0.225 mM dNTP, 0.8 μM primers, 100 ng primers, 1 unit AmpliTaq Gold™(Applied Biosystems), 1X Taq buffer.

607–612del/insTG (EIF2B2), A638G (EIF2B2)

F: GGAAATTATGTGCTGGATATG

R: ACTTTATTCTCTCACCGTGGAT

 

P243L (EIF2B4)

F: ATGCTCAAGCTCCCTTTCAA

R: CTTCACAACTTACAAAGCCT

 

R374C (EIF2B4)

F: ATTCAAGCACCTGGCATGAT

R: CGCTGCACTCCATCCTTATC

PCR reaction: 95°C 12 min, 35 cycles (94°C 30 s, 55°C 30 s, 72°C 45 s), 72°C 10 min, 4°C.

T1393C (EIF2B4), T1465C (EIF2B4)

F: TGTCCTGTAAGTAGGGGACCTT

R: AAGGGGTTGTGAAGTCTGGA

 

G338A (EIF2B5), C583T (EIF2B5)

F: GAGAAGGACTGTGAGTGCTGA

R: GCCTTCTAAGGGGACAATAAC

 
  1. F: forward primer, R: reverse primer