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Table 1 Oligonucleotides used for EBV QPCR, beta-globin detection, construction of plasmid and PCR thermal cycling conditions.

From: Measurement of Epstein-Barr virus DNA load using a novel quantification standard containing two EBV DNA targets and SYBR Green I dye

Target Primer Name Oligonucleotide Sequence 5'-3' Amplicon Length GenBank Accession (position) Reference Optimised PCR Thermal Cycling Conditions
EBNA-1 QP1 GCC GGT GTG TTC GTA TAT GG 213 bp AJ507799
(97174-97386)
Stevens et al, 1999 95°C initial denaturation for 10 mins; 58°C annealing
  QP2 CAA AAC CTC AGC AAA TAT ATG AG     
BHRF-1 EA-1F GGA GAT ACT GTT AGC CCT G 208 bp AJ507799
(42105-42312)
Custom 98°C initial denaturation for 13 mins; 60°C annealing
  EA-2R GTG TGT TAT AAA TCT GTT CCA AG     
Plasmid construct (randomised primers in bold) EA-F CTA TAT GTC TGC TTA CTC CGG CG /G GAG ATA CTG TTA GCC CTG 554 bp N/A Custom 95°C initial denaturation for 10 mins; 55°C annealing
  EB-R CGC CGG AGT AAG CAG ACA TAT AG /CAA AAC CTC AGC AAA TAT ATG AG     95°C initial denaturation for 10 mins; 55°C annealing
Beta-Globin TAL57 BG-1F TAG CAA CCT CAA ACA GAC ACC A 247 bp EU760960
(171-417)
Custom 95°C initial denaturation for 10 mins; 61°C annealing
  BG-1R CAG CCT AAG GGT GGG AAA AT     
  1. Abbreviations: EBNA-1, Epstein-Barr virus nuclear antigen-1; BHRF-1, BamHI fragment H rightward open reading frame-1; mins, minutes