Table 1

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

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


Abbreviations: EBNA-1, Epstein-Barr virus nuclear antigen-1; BHRF-1, BamHI fragment H rightward open reading frame-1; mins, minutes

Lay et al. Virology Journal 2010 7:252   doi:10.1186/1743-422X-7-252

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