Validated primer pairs specific to North America H3, H5, and H7 
avian influenza subtypes.  Predicted Tm and optimum thermodynamic 
alignments with primer complements and the three plasmids used 
in validation are included.  Primers were designed by running 
PrimerHunter with the following parameters:
- Primer length between 20 and 25
- Amplicon length between 75 and 200
- GC content between 25% and 75%
- Maximum mononucleotide repeat of 5
- 3'-end match mask M=11
- No required 3' GC Clamp
- Primer concentration of 0.8uM
- Salt concentration of 50mM
- Tmintarget = 35°C
- Tmaxnontarget = 45°C
- Delta between min with targets and max with non targets = 7°C

(Plasmid sequences were NOT used as targets in the design of the 
primers.)


### Pair H3-A ###

H3-A-Forward $GGATCTTATGGATTTCCTTTGCCAT$
Complement   $CCTAGAATACCTAAAGGAAACGGTA$ Tm: 58.17 
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H3-A-Forward $GGATCTTATGGATTTCCTTTGCCAT$
H3 Plasmid   ACCTAGAATACCTAAAGGAAACGGTAT Tm: 58.73
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H3-A-Forward $GGATCTTATGGATTTCCTTTGCCAT$
H5 Plasmid   AAGGTGTTACCAGTAAGGAGATGGTAA Tm: 10.00
             .....M.MMMM...MMMMM.M.MMMM.

H3-A-Forward $GGATCTTATGGATTTCCTTTGCCAT$
H7 Plasmid   CCCTA----ACCAAAAGTAAACG-TAT Tm: 13.76
             .MMMM....MMM.MMMM.MMMMM.MM.


H3-A-Reverse $TTTGGCAGGCCCACATAATGAACCC$
Complement   $AAACCGTCCGGGTGTATTACTTGGG$ Tm: 64.48 
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H3-A-Reverse $TTTGGCAGGCCCACATAATGAACCC$
H3 Plasmid   GAAACCGTCCGGGTGTATTACTTGGGG Tm: 63.69
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H3-A-Reverse $TTTGGCAGGCCCACATAATGAACCC$
H5 Plasmid   GGTACTAACCGGGT-----TCTTGGGA Tm: 19.98
             ...MM...MMMMMM......MMMMMM.

H3-A-Reverse $TTTGGCAGGCCC--ACATAATGAACCC$
H7 Plasmid   AAAACCTACCGGACTTGTA-----TGGGT Tm: 14.77
             .MMMMM..MMMM...MMMM.....MMMM.


### Pair H3-B ###

H3-B-Forward $GCTTCAAAATATACCACAAATG$
Complement   $CGAAGTTTTATATGGTGTTTAC$ Tm: 51.68 
             .MMMMMMMMMMMMMMMMMMMMMM.

H3-B-Forward $GCTTCAAAATATACCACAAATG$
H3 Plasmid   ACGAAGTTTTATATGGTGTTTACA Tm: 53.38
             .MMMMMMMMMMMMMMMMMMMMMM.

H3-B-Forward $GCTTCAAAATATACCACAAATG$
H5 Plasmid   AAGGTAATATCCGTGGGGTTTGCA Tm: 6.10
             ..M....M.M...MMM.MMMM.M.

H3-B-Forward $GCTTCAAAATATACCACAAATG$
H7 Plasmid   TCG-AGTTTTA--TGGTCGTTAGT Tm: 6.46
             .MM.MMMMMMM..MMMM..MMM..


H3-B-Reverse $AGGAAATCCATAAGATCCAGTCTTT$
Complement   $TCCTTTAGGTATTCTAGGTCAGAAA$ Tm: 56.36 
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H3-B-Reverse $AGGAAATCCATAAGATCCAGTCTTT$
H3 Plasmid   TTCCTTTAGGTATTCTAGGTTAGAAAC Tm: 52.18
             .MMMMMMMMMMMMMMMMMMM.MMMMM.

H3-B-Reverse $AGGAAATCCATAAGATCCAGTCTTT$
H5 Plasmid   TAAATTAGTACCTTTAGGTCCAAGAAAG Tm: 11.48
             $AGGAAATCCATAGATCCAG--TCTTT$
             ....MM......M.MMMMMM..MMMMM.

H3-B-Reverse $AGGAAATCCATAAGATCCAGTCTTT$
H7 Plasmid   TTCGATTTGGTATTCTAATACAGAAAC Tm: 26.29
             .MM..MM.MMMMMMMMM...MMMMMM.


### Pair H3-C ### 

H3-C-Forward $ACAGCACTGCAACACTGTGCTTGGG$
Complement   $TGTCGTGACGTTGTGACACGAACCC$ Tm: 66.89 
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H3-C-Forward $ACAGCACTGCAACACTGTGCTTGGG$
H3 Plasmid   TTGTCGCGACGTTGTGACACGAACCCC Tm: 62.51
             .MMMMM.MMMMMMMMMMMMMMMMMMM.

H3-C-Forward $ACAGCACTGCAACACTGTGCTTGGG$
H5 Plasmid   CTTTCCCCTGGTTTAGACGC-AACCCA Tm: 17.96
             .M.MM.....MMM..MMM.M.MMMMM.

H3-C-Forward $ACAGCACTGCAACACTGTGCTTGGG$
H7 Plasmid   GATTTCCTCTGTTTTATACAGAACCCG Tm: 10.50
             ...M......MMM.M.....MMMMMM.


H3-C-Reverse $TGTTGCATATTTTCCCTGTTGAGGA$
Complement   $ACAACGTATAAAAGGGACAACTCCT$ Tm: 59.52 
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H3-C-Reverse $TGTTGCATATTTTCCCTGTTGAGGA$
H3 Plasmid   AACAACGTATAAAAGGGACAACTTCTC Tm: 53.80
             .MMMMMMMMMMMMMMMMMMMMMM.MM.

H3-C-Reverse $TGTTGCATATTTTCCCTGTTGAGGA$
H5 Plasmid   GTACGGCTGGGGTGGGGACTACTCCGT Tm: 22.42
             .......M......MMMMM.MMMMM..

H3-C-Reverse $TGTTGCATATTTTCCCTGTTGAGGA$
H7 Plasmid   ACCCATGTATAAAAG----AACTATAA Tm: 8.53
             ..M...MMMMMMMMM....MMMM....


### Pair H5-A ###
 
H5-A-Forward $TCTGGATTTCCATGATTCAAATGTC$
Complement   $AGACCTAAAGGTACTAAGTTTACAG$ Tm: 56.64 
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H5-A-Forward $TCTGGATTTCCATGATTCAAATGTC$
H3 Plasmid   TCTTCCTAAGTGTAC-CTGTCCTCAGT Tm: 2.50
             ....MMMMM..MMMM...MM...MMM.

H5-A-Forward $TCTGGATTTCCATGATTCAAATGTC$
H5 Plasmid   TGGACCTAAAGGTACTAA-TTTACAGT Tm: 44.94
             ..MMMMMMMMMMMMMMMM.MMMMMMM.

H5-A-Forward $TCTGGATTTCCATGATTCAAATGTC$
H7 Plasmid   AAGTCCGGTAGGT------TTTACGGT Tm: 5.57
             .MM.MM...MMMM......MMMMM.M.


H5-A-Reverse $ACGTTCCATTTCTCACACTTTCCAT$
Complement   $TGCAAGGTAAAGAGTGTGAAAGGTA$ Tm: 59.75
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H5-A-Reverse $ACGTTCCATTTCTCACACTTTCCAT$
H3 Plasmid   TAACAAGGTAAACCGTGGC----GTAC Tm: 4.78
             ...MMMMMMMMM..MMM......MMM.

H5-A-Reverse $ACGTTCCATTTCTCACACTTTCCAT$
H5 Plasmid   TATGAAGGTAAAGAGTGTGAAAGGTAT Tm: 54.77
             ....MMMMMMMMMMMMMMMMMMMMMM.

H5-A-Reverse $ACGTTCCATTTCTCACACTTTCCAT$
H7 Plasmid   AGACCTTGTAAAGAGT----AAGGGAC Tm: 11.75
             ...M...MMMMMMMMM....MMMM.M.


### Pair H5-B ###
 
H5-B-Forward $ATGGAATTTTTCTGGACAATACTAA$
Complement   $TACCTTAAAAAGACCTGTTATGATT$ Tm: 53.60 
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H5-B-Forward $ATGGAATTTTTCTGGACAATACTAA$
H3 Plasmid   CTATTCGTAGATAACCTGTTAT----- Tm: 1.48
             .MM.....M.M..MMMMMMMMM.....

H5-B-Forward $ATGGAATTTTTCTGGACAATACTAA$
H5 Plasmid   ACCCTTAAAAAAGACCTGTTATGATTC Tm: 43.89
             ...M.M.MMMMMMMMMMMMMMMMMMM.

H5-B-Forward $ATGGAATTTTTCTGGACAATACTAA$
H7 Plasmid   TTGTCTAGACCCTGTTACACCTGA--- Tm: -4.78
             .M..M.MMMMM.MMMMM....MMM...


H5-B-Reverse $TGTATGCATATTCTGGAGCTATAAA$
Complement   $ACATACGTATAAGACCTCGATATTT$ Tm: 54.64
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H5-B-Reverse $TGT-ATGCATATTCTGGAGCTATAAA$
H3 Plasmid   CTCACTACGTATCAGACCATGTAGTATA Tm: 9.70
             ..MM.MMMMMMM.MMMMM..M...M.M.

H5-B-Reverse $TGTATGCATATTCTGGAGCTATAAA$
H5 Plasmid   AACATGCGTATAAGTCCTCGATATTTT Tm: 47.25
             .MMMM.MMMMMMMM.MMMMMMMMMMM.

H5-B-Reverse $TGTATGCAT-ATTCTGGA--GCTATAAA$
H7 Plasmid   TTTCCTCGTAGTGAGACCTAAGGAT----- Tm: 0.58
             ......MMMM.M.MMMMMM...MMM.....


### Pair H5-C ###
 
H5-C-Forward $GACGGTCACACATGCTCAGGACATA$
Complement   $CTGCCAGTGTGTACGAGTCCTGTAT$ Tm: 63.25 
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H5-C-Forward $GACGGTCACACATGCTCAGGACATA$
H3 Plasmid   TTTGCCC-TGGATACTAGTGCTGTATA Tm: 16.93
             ..MMMM..MM..MMM.MMM.MMMMMM.

H5-C-Forward $GACGGTCACACATGCTCAGGACATA$
H5 Plasmid   CCTGCCCCTGTGTACGAGTCCTATATG Tm: 45.65
             .MMMMM..MMMMMMMMMMMMMM.MMM.

H5-C-Forward $GACGGTCACACATGCTCAGGACATA$
H7 Plasmid   GGTACTTACCCATACAAGTCCGGTAGG Tm: 4.36
             ..M.M.......MMM.MMMMM.MMM..


H5-C-Reverse $CACTGCAATCCTTCAGAATGAGGGG$
Complement   $GTGACGTTAGGAAGTCTTACTCCCC$ Tm: 62.52
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H5-C-Reverse $CACTGCA---ATCCTTCAGAATGAGGGG$
H3 Plasmid   TAAGACGTCTATAGGTAGTGTGACCGCCGG Tm: 17.38
             ...MMMMM...MMMM.MMM.M.MM..MM..

H5-C-Reverse $CACTGCAATCCTTCAGAATGAGGGG$
H5 Plasmid   CATGGACTTAGGAAGTCTTACTCCCCA Tm: 50.57
             ..MM...MMMMMMMMMMMMMMMMMMM.

H5-C-Reverse $CACTGCAATCCTTCAGAATGAGGGG$
H7 Plasmid   TAAGGGACCATCCGGTCTTCCTCCCGA Tm: 21.02
             ...M.....M....MMMMM.MMMMM..


### Pair H7-A ###
 
H7-A-Forward $ACCCAGTGAAATTGAGTAGTGGATA$
Complement   $TGGGTCACTTTAACTCATCACCTAT$ Tm: 58.07 
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H7-A-Forward $ACCCAGTGAAATTGAGTAGTGGATA$
H3 Plasmid   ATGGGTCAC---AACTTA-CACTGATA Tm: 16.80
             .MMMMMMMM...MMMM.M.MMM..MM.

H7-A-Forward $ACCCAGTGAAATTGAGTAGTGGATA$
H5 Plasmid   CTCTTTCACTTAAACTCAT-ACCATGG Tm: 18.03
             .M...MMMMMM.MMMMMMM.MMM....

H7-A-Forward $ACCCAGTGAAATTGAGTAGTGGATA$
H7 Plasmid   CTGGGTCACTTCAACTCATCACCTATG Tm: 52.61
             .MMMMMMMMMM.MMMMMMMMMMMMMM.


H7-A-Reverse $AGTGCACCGCATGTTTCCATTCTTT$
Complement   $TCACGTGGCGTACAAAGGTAAGAAA$ Tm: 62.69 
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H7-A-Reverse $AGTGCACCGCATGTTTCCATTCTTT$
H3 Plasmid   ACAACGTGGATTACAACGG--AGAAAC Tm: 25.17
             ...MMMMMM..MMMMM.MM..MMMMM.

H7-A-Reverse $AGTGCACCGCATGTTTCCATTCTTT$
H5 Plasmid   GACGCGTGGC$---------------- Tm: 18.77
             ..M.MMMMMM.................

H7-A-Reverse $AGTGCACCGCATGTTTCCATTCTTT$
H7 Plasmid   ATCACGTGGCGTACAAAGGTAAGAAAT Tm: 63.09
             .MMMMMMMMMMMMMMMMMMMMMMMMM.


### Pair H7-B ###
 
H7-B-Forward $ATAGAATACAGATAGACCCAGTGAA$
Complement   $TATCTTATGTCTATCTGGGTCACTT$ Tm: 55.23 
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H7-B-Forward $ATAGAATACAGATAGACCCAGTGAA$
H3 Plasmid   TTTAGACCTTCATGAATGGGTCACAAC Tm: 16.52
             .M......M...M...MMMMMMMM...

H7-B-Forward $ATAGAATACAGATA-----GACCCA-GTGAA$
H5 Plasmid   TTCTTGGATATCTATTCCAGCTGGGTGCACTCT Tm: 5.79
             .M.M...MM.MMMMM.....MMMMMM.MMMM..

H7-B-Forward $ATAGAATACAGATAGACCCAGTGAA$
H7 Plasmid   TTATCTTATGTCTATCTGGGTCACTTC Tm: 55.21
             .MMMMMMMMMMMMMMMMMMMMMMMMM.


H7-B-Reverse $AAACTATATACAAATAGTGCACCGC$
Complement   $TTTGATATATGTTTATCACGTGGCG$ Tm: 56.58 
             .MMMMMMMMMMMMMMMMMMMMMMMMM.

H7-B-Reverse $AAACTATATACAAATAGTGCACCGC$
H3 Plasmid   AAGTGATAACAAGGTAAACCGTGGCGT Tm: 28.79
             ...MMMMM......MM...MMMMMMM.

H7-B-Reverse $AAACTATATACAAATAGTGCACCGC$
H5 Plasmid   GCTCAGCGCAGGTTCGACGCGTGGC$- Tm: 19.78
             ..M......M.MMM...M.MMMMMM..

H7-B-Reverse $AAACTATATACAAATAGTGCACCGC$
H7 Plasmid   GTTTGATATATGTTTATCACGTGGCGT Tm: 57.24
             .MMMMMMMMMMMMMMMMMMMMMMMMM.


### Pair H7-C ###
 
H7-C-Forward $AAGATTAGGTTCTTCCTTCTATGC$
Complement   $TTCTAATCCAAGAAGGAAGATACG$ Tm: 55.2 
             .MMMMMMMMMMMMMMMMMMMMMMMM.

H7-C-Forward $AAGAT-TAGGTTCTTCCTTCTATGC$
H3 Plasmid   GTTCTTGATCCCAGAACAAAGCCACGT Tm: 6.85
             .MMMM..MMMM.MMMM..MMM..MMM.

H7-C-Forward $AAGATTAGGTTCTTCCTTCTATGC$
H5 Plasmid   ACCCACGATATTTGAGGGTCATACGG Tm: -2.47
             ...M..........MMM...MMMMM.

H7-C-Forward $AAGATTAGGTTCTTCCTTCTATGC$
H7 Plasmid   GTTCTAGTCCGAGAAGAAAGATACGT Tm: 39.40
             .MMMMM.MMM.MMMMM.MMMMMMMM.


H7-C-Reverse $AACTTGTTTCCACTTCCATAG$
Complement   $TTGAACAAAGGTGAAGGTATC$ Tm: 53.44 
             .MMMMMMMMMMMMMMMMMMMMM.

H7-C-Reverse $AACTTGTTTCCACTTCCATAG$
H3 Plasmid   AGTGAGCTAAGTTGAAGGGTAAT Tm: 4.03
             ..MMM.M.MMM.MMMMMM.....

H7-C-Reverse $AACTTGTTTCCA--CTTCCATAG$
H5 Plasmid   GATATTTTAAGGTATGAAGGT-TCC Tm: 9.82
             ..M.....MMMMM..MMMMMM.MM.

H7-C-Reverse $AACTTGTTTCCACTTCCATAG$
H7 Plasmid   GTTGAACAAAGGTGAAGGTATCT Tm: 54.44 
             .MMMMMMMMMMMMMMMMMMMMM.