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Oligonucleotide primers used in the WRLFMD
for RT-PCR and cycle sequencing of foot-and-mouth disease viruses.
Primer Primer (5' 6 3') Location__________________
designation_____sequence________________________Gene___Nucleotides___Use____
Positive sense primers
ARS4 ACCAACCTCCTTGATGTGGCT
VP3 124-145
PCR
A-1C562 TACCAAATTACACACGGGAA
VP3 562-581
PCR
C-1C536 TACAGGGATGGGTCTGTGTGTACC
VP3
536-559 PCR
As1-1C505 TACACTGCTTCTGACGTGGC
VP3 505-524
PCR
SAT-1D209F CCACATACTACTTTTGTGACCTGGA
VP1 209-234
PCR
Negative sense primers
NK61
GACATGTCCTCCTGCATCTG
2B 58-77
RT+PCR
SAT-2B208R ACAGCGGCCATGCACGACAG
2B 208-227
RT+PCR
NK72
GAAGGGCCCAGGGTTGGACTC
2A/2B 34-48; 1-6 Sequencing
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Table 2. Primer combinations used in the WRLFMD to RT-PCR amplify
foot-and-mouth disease viruses.
Primers______________________________________
Serotype__ RT*_________Forward_______Reverse________Product length
(bp)__
O NK61 ARS4 NK61 1301
A NK61 A-1C562 NK61 863 to 866
C NK61 C-1C536 NK61 877 to 883
Asia 1 NK61 As1-1C505 NK61 908 to 914
SAT 1 SAT-2B208R SAT-1D209F SAT-2B-208R 731
SAT 2 SAT-2B208R SAT-1D209F SAT-2B-208R 716
SAT 3 SAT-2B208R SAT-1D209F SAT-2B-208R 725
* reverse transcription; alternatively random hexamers may be
used.
For the O/A/C/Asia 1 primers: Knowles, N.J. and Samual, A.R.,
(1994)
For the SAT primers: N.J. Knowles, unpublished data
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