SEQUENCE LISTING
<110> AMIRUL ISLAM
<120> A METHOD TO AMPLIFY AND DETECT TARGET NUCLEIC ACIDS WITH VERY HIGH
SPECIFICITY AND SENSITIVITY
<130> ASA PP-3330/AI
<160> 27
<170> Patent In version 3.5[SI1]

<210> 1
<211> 21
<212> DNA
<213> E.coli unlabelled forward primer
<400> 1
gcacaaatgc tgacccatat t                                                             	21

<210> 2
<211> 22
<212> DNA
<213> E.coli
<220> BHQ1 DYE
<221> modified base
<222> 18
<223> Blackhole quencher 1 labelled reverse primer
<400> 2
tcacattcgg taaaccgtag aa 					                          22

<210> 3
<211> 120
<212> DNA
<213> E. coli target DNA for forward primer seq 1, reverse primer seq 4, seq 2 & probe seq 5, seq 6
<400> 3
tggcacaaat gctgacccat attgcgggtg ataagccagt gaccattctg accgcgacct ccggtgatac cggagcggca gtggctcatg ctttctacgg tttaccgaat gtgaaagtgg 					            120 

<210> 4
<211> 22
<212> DNA
<213> E. coli
<220> BHQ1 DYE
<221> modified base
<222> 7
<223> Blackhole quencher 1 labelled reverse primer
<400> 4
tcacattcgg taaaccgtag aa 					                            22

<210> 5
<211> 24
<212> DNA
<213> E. coli
<220> FLUORESCEIN DYE
<221> modified base
<222> 24
<223> Fluorescein dye labelled probe
<400> 5
gggtgataag ccagtgacca ttct 					                             24

<210> 6
<211> 24
<212> DNA
<213> E.coli
<220> FLUORESCEIN DYE
<221> modified base
<222> 21
<223> Fluorescein labelled probe
<400> 6
gggtgataag ccagtgacca ttct 					                             24

<210> 7 
<211> 21
<212> DNA
<213> E. coli
<220> FLUORESCEIN DYE
<221> modified base
<222> 3
<223> Fluorescein labelled forward primer
<400> 7
tctcggcgtt tattggtgat g 						                     21

<210> 8  
<211> 21
<212> DNA
<213> E.coli
<220> FLUORESCEIN DYE
<221> modified base
<222> 13
<223> Fluorescein labelled forward primer
<400> 8
tctcggcgtt tattggtgat g                                                                       21

<210> 9 
<211> 21
<212> DNA
<213> E. coli
<220> FLUORESCEIN DYE
<221> modified base
<222> 10
<223> Fluorescein labelled forward primer
<400> 9
tctcggcgtt tattggtgat g                                                                       21

<210> 10  
<211> 20
<212> DNA
<213> E.coli
<220> BHQ1 DYE
<221> modified_base
<222> 16
<223> Blackhole quencher 1 labelled (on base 16) reverse primer
<400> 10
gacaaccgac atcgctttc                                                                          20

<210> 11 
<211> 24
<212> DNA
<213> E.coli
<220> FLUORESCEIN DYE
<221> modified base
<222> 1
<223> MODIFICATION:  FLUORESCIN DYE ON BASE 1 
<400> 11
gggtgataag ccagtgacca ttct                                                                    24

<210> 12 
<211> 21
<212> DNA
<213> E coli Forward primer sequence
<220> FLUORESCEIN DYE
<221> modified base
<222> 1
<223> FLUORESCEIN DYE on Base 1 
<400> 12
tctcggcgtt tattggtgat g                                                                       21

<210> 13
<211> 20
<212> DNA
<213> E. coli forward primer
<400> 13
gataagctgc cgtcagaacc                                                                         20

<210> 14
<211> 20
<212> DNA
<213> E.coli reverse primer
<400> 14
aacaggcact ggagcctaag                                                                         20

<210> 15
<211> 21
<212> DNA
<213> E. coli
<220> FLUORESCEIN DYE & NFQ DYE
<221> modified base
<222> 1 & 21
<223> Fluorescein at 5 end on Base 1 and quencher NFQ at 3 end on base 21
<400> 15
tccagggtca tcgccactgg a 						                      21

<210> 16
<211> 46
<212> DNA
<213> Artificial sequence
<220>
<223> Synthetic target sequence for hybridizing sequence id 5 (Probe) and sequence id 6 (Probe)  
and sequence id 4 (primer) to generate different separations between Fluorescein and Blackhole Quencher 1
<400> 16
ttctacggtt taccgaatgt gaagaatggt cactggctta tcaccc                         		      46
<210> 17
<211> 48
<212> DNA
<213> Artificial sequence
<220>
<223> Synthetic target sequence for hybridizing sequence id 5 (Probe) and sequence id 6 (Probe)  
and sequence id 4 (primer) to generate different separations between Fluorescein and Blackhole Quencher 1
<400> 17
ttctacggtt taccgaatgt gataagaatg gtcactggct tatcaccc					      48

<210> 18
<211> 45
<212> DNA
<213> Artificial sequence
<220>
<223> Synthetic target sequence for hybridizing sequence id 7 (Primer) and sequence id 8 (Primer)  
and sequence id 9 (primer) to measure enhancement of Fluorescein dye (FAM)
<400> 18
catcaccaat aaacgccgag aagaatggtc actggcttat caccc                         		      45
<210> 19
<211> 65
<212> DNA
<213> Artificial Sequence
<220>
<223> non-target primer sequence at 5 end and E coli threonine synthase gene
sequence at its 3 end
<400> 19
agatgtccgt gctaccagag  actctcgggc ggtcgcttta tggcacaaat gctgacccat attgc                      65

<210> 20
<211> 60
<212> DNA
<213> Artificial Sequence
<220>
<223> E coli threonine synthase gene sequence at 5 end and a non-target primer
sequence at its 3 end
<400> 20
gggtgataag ccagtgacca ttctgaccgc gacctagaca ccggtataac gacctgccca                             60

<210> 21
<211> 27
<212> DNA
<213> Artificial Sequence
<220> Blackhole Quencher 1
<221> modified_base
<222> 21
<223> Blackhole Quencher1 labeled (on base 23) first non-target primer
<400> 21
agatgtccgt gctaccagag actctcg                                                                 27

<210> 22
<211> 25
<212> DNA
<213> Artificial Sequence
<220> Fluorescein dye
<221> modified_base
<222> 15
<223> Fluorescein labeled second non-target primer (ON BASE 15)
<400> 22
tgggcaggtc gttataccgg tgact    								     25                                                                      25

<210> 23
<211> 20
<212> DNA
<213> E.coli unlabelled forward primer
<400> 23
gataagctgc cgtcagaacc                                                                        20 

<210> 24
<211> 20
<212> DNA
<213> E.coli
<220> FLUORESCEIN DYE
<221> Modified_base
<222> 17
<223> Fluorescein labeled (on base 17)reverse primer
<400> 24
aacaggcact ggagcctaag                                                                        20

<210> 25
<211> 30
<212> DNA
<213> E. coli
<220> FLUORESCEIN DYE
<221> Modified_base
<222> 27
<223> Fluorescein labeled probe on base 27
<400> 25
ccagtggcga tgaccctgga aaagaatatg                                                             30

<210> 26
<211> 120
<212> DNA
<213> E. coli target DNA for forward primer seq 7, seq 8, seq9 and reverse primer seq 10
<400> 26
gatcctctcg gcgtttattg gtgatgaaat cccacaggaa atcctggaag agcgcgtgcg 
cgcggcgttt gccttcccgg ctccggtcgc caatgttgaa agcgatgtcg gttgtctgga         		     120

<210> 27
<211> 73
<212> DNA
<213> E. coli target DNA for amplification with non-target primers seq 21, and seq 22 
<400> 27
ggcggtcgct ttatggcaca aatgctgacc catattgcgg gtgataagcc agtgaccatt ctgaccgcga cct             73

<210> 28
<211> 145
<212> DNA
<213> E. coli target DNA for primers seq 23, seq 24 and probe 25  and primers seq 13, 14 and probe 15 for Taqman probe assay 
<400> 28
gataagctgc cgtcagaacc acgggaaaat atcgtttatc agtgctggga gcgtttttgc 
caggaactgg gtaagcaaat tccagtggcg atgaccctgg aaaagaatat gccgatcggt 
tcgggcttag gctccagtgc ctgtt                      					     145

