Table 1.

Intron spanning primers used in the expression analysis.

Gene	CADRE	Forward	pMol	Reverse	pMol
Asp f 1	AFUA_5G02330	ACGCTCGTGCG*ACCTGGACATGC	40	GCCGTCGGAAAGAGGTGCGTG	20
Asp f 2	AFUA_4G09580	CTGTGCTTTGGAAG*GCTGGGGCGGCCAC	40	GTCTCCATGTGCTCCCAGGGC	40
Asp f 3	AFUA_6G02280	GGGACGACATT*CTCTTCCTCTCCGAC	40	CGCTCGAGAACTCGAGGTGGTTC	40
Asp f 4	AFUA_2G03830	CAGCTCTTCCCACTCCGACAG	40	CTGGGTTCGGTCCTGCCAC	40
Asp f 5	AFUA_8G07080	TACTCACGGTC*TTTCCAACCGAC	40	GCTTCAGACGGATGGCCGTC	40
Asp f 6	AFUA_1G14550	ACTACCTTCAG*TACTTGAACGAC	100	GTACACGTTCATGAATGGGTG	40
Asp f 7	AFUA_4G06670	GCTCCTATCTTCAAGTCCCT	40	CCACACTACGTCCACTTCAC	40
Asp f 8	AFUA_2G10100	ACCTCCAGGAGCTCATCGCCGAG	20	CTCCTCCTTCTTCTCCTCAG	40
Asp f 9	AFUA_1G16190	GAGGTTGACTGG*GAAGTATTG	40	GAAAGTCTCCTGAGGAGTG	10
Asp f 10	AFUA_5G13300	GCGGCATTGCTG*ACACCGGC	40	GCAGGGGAAGACATAACCACCG	40
Asp f 11	AFUA_2G03720	GGTCCTAACAC*CAACGGC	40	GAGCTTCGATCTCCTTGAC	40
Asp f 12	AFUA_5G04170	TGACCAAGGCT*GATTTGATC	40	CAACAAGGTAAGCAGAGTAG	40
Asp f 13	AFUA_4G11800	GAGCGCAGAC*GTTGCCCATG	40	CCTTGTGGGAAATGCTGCCCAG	40
Asp f 17	AFUA_4G03240	ACCATCAACTCCGGTGTCGAC	10	CTTGGAGATGAGGTCGTCG	40
Asp f 18	AFUA_5G09210	CTCCCAAC*CTCCTTGCCTG	40	CTCGGCCTTGTGAACTAG	40
Asp f 22	AFUA_6G06770	CATGATCGTCCCTGA*CTCCGC	40	CACCCTCGTCACCAACGTTG	40
Asp f 23	AFUA_2G11850	GCAGATTACTCC*CATGGGTG	40	GTACAGGGTCTTGCGCAG	40
Actin	AFUA_6G04740	TCATCATGCGCGACAGC	10	CAATCATGATA*CCATGGTGAC	40
β-tubulin	AFUA_1G10910	CGACAACGAG*GCTCTGTACG	40	CAACTTGCGCAGATCAGAGTTGAG	40
GpdA	AFUA_5G01970	GGCGAGCTCAAGAACATCCTCGGCTA	20	CTTGGCGATGTAGGCGATAAGGTCGA	20
FksA	AFUA_6G12400	GCTGCGCCCAAG*TCGCCAAATC	40	GAACAACAAGTGGGGCAATG	20

As part of the initial work up for these experiments primer concentrations were optimized so that all allergens could be analysed using a single annealing temperature in the PCR. Primer levels used are indicated. Primers used to establish the most useful comparator are also included.

pMol = pMol primer used per 25 μl reaction mix; * - intron site