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Table 1 Intron spanning primers used in the expression analysis.

From: Aspergillus fumigatus allergen expression is coordinately regulated in response to hydrogen peroxide and cyclic AMP

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

  1. 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.
  2. pMol = pMol primer used per 25 μl reaction mix; * - intron site