                         SEQUENCE LISTING

<110>  Biofidelity Ltd
 
<120>  IMPROVED POLYNUCLEOTIDE SEQUENCE DETECTION METHOD

<130>  P32887WO1

<150>  2020539.9
<151>  2020-12-23

<150>  2101176.2
<151>  2021-01-28

<160>  30    

<170>  PatentIn version 3.5

<210>  1
<211>  37
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Splint oligonucleotide seq

<400>  1
acagtttcgg agtacgaaca tcggtgcgtt tcgcaag                                37


<210>  2
<211>  82
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Probe oligonucleotide seq


<220>
<221>  5Phos
<222>  (1)..(1)
<223>  5' phosphorylation

<220>
<221>  Phosphorothioate-bond
<222>  (1)..(4)
<223>  Phosphorothioate bonds between bases 1&2, 2&3, 3&4

<400>  2
atgttcgatg agctttgaca atacttgacg attcatgccg tacatgcagc tatgcagata       60

taggatgttg cgaaacgcac cg                                                82


<210>  3
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer mix 1 Fwd

<400>  3
ggcatgaatc gtcaagtatt                                                   20


<210>  4
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer mix 1 Rev

<400>  4
atgcagatat aggatgttgc                                                   20


<210>  5
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Labeled probe seq


<220>
<221>  5Cy5
<222>  (1)..(1)
<223>  Cy5 on 5' end

<220>
<221>  TAO
<222>  (9)..(10)
<223>  TAO fluorescent quencher between bases 9&10

<220>
<221>  3IAbRQSp
<222>  (20)..(20)
<223>  Iowa Black RQ fluorescence quencher on 3' end

<400>  5
cgcaccgatg ttcgatgagc                                                   20


<210>  6
<211>  51
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Capture oligonucleotide seq


<220>
<221>  5biotin
<222>  (1)..(1)
<223>  Biotin on 5' end

<400>  6
tttttttttt tttttttttt atctttttga attcagtttc cttcatctag g                51


<210>  7
<211>  97
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Probe seq


<220>
<221>  5Phos
<222>  (1)..(1)
<223>  Phosphorylated 5' end

<220>
<221>  Phosphorothioate-bond
<222>  (1)..(4)
<223>  Phosphorothioate bonds between bases 1&2, 2&3, 3&4

<400>  7
atgttcgatg agctttgaca atacttgacg attcatgccg tacatgcagc tatgcagata       60

taggatgttg cgaaacgcac cggaggccag cactttg                                97


<210>  8
<211>  70
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Wildtype (WT) oligonucleotide seq

<400>  8
aaggaaactg aattcaaaaa gatcaaagtg ctgggctccg gtgcgttcgg cacggtgtat       60

aaggtaaggt                                                              70


<210>  9
<211>  70
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Mutant oligonucleotide seq

<400>  9
aaggaaactg aattcaaaaa gatcaaagtg ctggcctccg gtgcgttcgg cacggtgtat       60

aaggtaaggt                                                              70


<210>  10
<211>  77
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Probe seq


<220>
<221>  5Phos
<222>  (1)..(1)
<223>  Phosphorylated 5' end

<220>
<221>  Phosphorothioate-bond
<222>  (1)..(1)
<223>  Phosphorothioate bonds between bases 1&2, 2&3, 3&4

<400>  10
atgttcgatg agctttgaca atacttgaca tgcgcagata taggatgttg cgaaacgcac       60

cggaggccag cactttg                                                      77


<210>  11
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer mix 1 Fwd


<220>
<221>  Phosphorothioate-bonds
<222>  (1)..(20)
<223>  Phosphorothioate bonds between bases 1&2, 2&3, 17&18, 18&19, 
       19&20

<400>  11
tcgcaacatc ctatatctgc                                                   20


<210>  12
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer mix 1 Rev


<220>
<221>  Phosphorothioate-bonds
<222>  (1)..(10)
<223>  Phosphorothioate bonds between bases 1&2, 2&3, 17&18, 18&19, 
       19&20

<400>  12
tgagctttga caatacttga                                                   20


<210>  13
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer mix 2 Fwd


<220>
<221>  Phosphorothioate-bonds
<222>  (18)..(21)
<223>  Phosphorothioate bonds between bases 18&19, 19&20, 20&21

<400>  13
acatcggtgc gtttcgcaac a                                                 21


<210>  14
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer mix 2 Rev


<220>
<221>  Phosphorothioate-bonds
<222>  (1)..(20)
<223>  Phosphorothioate bonds between bases 1&2, 2&3, 17&18, 18&19, 
       19&20

<400>  14
tgagctttga caatacttga                                                   20


<210>  15
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer mix Fwd

<400>  15
tcgcaacatc ctatatctgc                                                   20


<210>  16
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer mix Rev

<400>  16
tgagctttga caatacttga                                                   20


<210>  17
<211>  70
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  WT oligo reverse complementary seq

<400>  17
accttacctt atacaccgtg ccgaacgcac cggagcccag cactttgatc tttttgaatt       60

cagtttcctt                                                              70


<210>  18
<211>  70
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Mutant oligo reverse complementary seq

<400>  18
accttacctt atacaccgtg ccgaacgcac cggaggccag cactttgatc tttttgaatt       60

cagtttcctt                                                              70


<210>  19
<211>  81
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  WT oligonucleotide seq

<400>  19
ctgctgggca tctgcctcac ctccaccgtg cagctcatca cgcagctcat gcccttcggc       60

tgcctcctgg actatgtccg g                                                 81


<210>  20
<211>  81
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Mutant oligonucleotide seq

<400>  20
ctgctgggca tctgcctcac ctccaccgtg cagctcatca tgcagctcat gcccttcggc       60

tgcctcctgg actatgtccg g                                                 81


<210>  21
<211>  74
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Probe A0 seq


<220>
<221>  5Phos
<222>  (1)..(1)
<223>  Phosphorylated 5' end

<220>
<221>  Phosphorothioate-bonds
<222>  (1)..(4)
<223>  Phosphorothioate bonds between bases 1&2, 2&3, 3&4

<400>  21
agctgcatct gagctttgac aatacttgag cacggcagat ataggatgtt gcgaagggca       60

tgagctgcat gatg                                                         74


<210>  22
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer mix Fwd


<220>
<221>  Phosphorothioate-bonds
<222>  (1)..(3)
<223>  Phosphorothioate bonds between bases 1&2, 2&3

<400>  22
tcgcaacatc ctatatctgc                                                   20


<210>  23
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer mix Rev

<400>  23
atgttgcgaa gggcatatgt                                                   20


<210>  24
<211>  49
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Capture oligonucleotide seq


<220>
<221>  5biotin
<222>  (1)..(1)
<223>  biotin on 5' end

<400>  24
tttttttttt tttttttttt aggcgggaga catatgggga gcccagtgg                   49


<210>  25
<211>  93
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Probe seq


<220>
<221>  5Phos
<222>  (1)..(1)
<223>  Phosphorylated 5' end

<220>
<221>  Phosphorothioate-bond
<222>  (1)..(2)
<223>  Phosphorothioate bond between residues 1&2

<220>
<221>  3SpC3
<222>  (93)..(93)
<223>  C3 spacer on 3' end

<400>  25
atgttcgatg agctttgaca atacttgatc gatgcagata taggatgttg cgaccatcac       60

gtatgccatc acgtatgctt cctggggaca ttt                                    93


<210>  26
<211>  90
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Unmethylated oligonucleotide seq

<400>  26
ctcagcgtac ccttgtcccc aggaagcata cgtgatggca tacgtgatgg ctggtgtggg       60

ctccccatat gtctcccgcc ttctgggcat                                        90


<210>  27
<211>  90
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Methylated oligonucleotide seq


<220>
<221>  iMe-dC
<222>  (20)..(20)
<223>  5-Methyl deoxycytidine

<400>  27
ctcagcgtac ccttgtcccc aggaagcata cgtgatggca tacgtgatgg ctggtgtggg       60

ctccccatat gtctcccgcc ttctgggcat                                        90


<210>  28
<211>  36
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Splint oligonucleotide seq

<400>  28
tgtcaaagct cagctatctg acgtgattcg caacaa                                 36


<210>  29
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer mix 1 Fwd


<220>
<221>  Phosphorothioate-bonds
<222>  (1)..(3)
<223>  Phosphorothioate bonds between bases 1&2, 2&3

<400>  29
tcgcaacatc ctatatctgc                                                   20


<210>  30
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer mix 1 Rev


<220>
<221>  Phosphorothioate-bonds
<222>  (1)..(3)
<223>  Phosphorothioate bonds between bases 1&2, 2&3

<400>  30
tgagctttga caatacttga                                                   20


