CN102230001A - Kit for detecting mutation of duchenne muscular dystrophy causing gene and use thereof - Google Patents
Kit for detecting mutation of duchenne muscular dystrophy causing gene and use thereof Download PDFInfo
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- CN102230001A CN102230001A CN2011101486070A CN201110148607A CN102230001A CN 102230001 A CN102230001 A CN 102230001A CN 2011101486070 A CN2011101486070 A CN 2011101486070A CN 201110148607 A CN201110148607 A CN 201110148607A CN 102230001 A CN102230001 A CN 102230001A
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Abstract
The invention discloses a kit for detecting if a duchenne muscular dystrophy (DMD) causing gene has a mutation. The kit comprises the following main components: (1) 79 pairs of primers for PCR amplification and sequencing of 1 to 79 extrons of the DMD causing gene; (2) a polymerase chain reaction (PCR) amplification reagent; (3) a PCR product purification reagent; and (4) a DNA sequencing reagent. The kit disclosed by the invention is used for detecting the mutation of a duchenne muscular dystrophy causing gene, can quickly and conveniently detect people or patient with the DMD causing gene, can be used for prenatal diagnosis, prevention of birth of affected children, and can lower the incidence rate of the DMD.
Description
Affiliated technical field
The invention belongs to biological technical field, be specifically related to detect the test kit whether the duchenne muscular dystrophy Disease-causing gene undergos mutation.
Background technology
Duchenne muscular dystrophy is the modal x linked recessive inherited disease of neural system, and the patient shows as that carrying out property of skeletal muscle is unable, atrophy, the false hypertrophy of shank gastrocnemius muscle, is divided into two types.Duchenne muscular dystrophy (Duchenne muscular dystrophy DMD) is more serious type, sickness rate be the life birth boy baby 1/3500. usually in about 20 years old because of respiratory insufficiency and (or) in heart failure dead.Becker ' s muscular dystrophy (Becker ' s muscular dystrophy, BMD) be a kind of symptom type light than DMD, sickness rate is 1/20000, one not influence patient's life-span.
Duchenne muscular dystrophy (Duehenne muscular dystrophy, DMD) be by due to the dystroplun transgenation of the encoding dystrophin that is positioned at Xp21 (dystrophin), the DMD patient that 60%-65% is arranged approximately is because due to the dystrophin gene extron disappearance, the 5%th, because due to the sudden change of dystrophin gene redundancy, the DMD morbidity of all the other 30%-35% is because due to the dystrophin point mutation (insertion of base substitution, transversion, disappearance, repetition or several bases etc.).
The pathogenesis of duchenne muscular dystrophy is fuzzy, does not still have effective methods of treatment so far.So detecting duchenne muscular dystrophy patient's DMD transgenation and finding out corresponding female carrier is the effective treatment measure that reduces or prevent the birth of duchenne muscular dystrophy infant.
Summary of the invention
The purpose of this invention is to provide a kind of test kit that adopts dna sequencing technology for detection duchenne muscular dystrophy Disease-causing gene DMD whether to suddenly change.
This detection kit mainly comprises following composition:
(1) DMD gene the 1st to the 79th exon district pcr amplification and sequencing primer are 79 pairs, and each sees Table 1 to primer sequence;
(2) conventional pcr amplification reagent is as dNTP, Taq archaeal dna polymerase etc.;
(3) conventional PCR product purification reagent is as SAP enzyme, ExoI enzyme etc.;
(4) conventional dna sequencing reagent is as BigDye mix, EDTA solution, ethanolic soln, HIDI solution etc.
The pcr amplification and the sequencing primer sequence in table 1 DMD gene 1-79 exon district
The use step of this test kit comprises:
(1) genomic dna of extraction sample;
(2) DMD gene PCR amplification:
Carry out pcr amplification at DMD gene 1-79 exon district according to table 1 design primer.Each reaction system is cumulative volume 10 μ l, comprises genomic dna (100ng/ μ l) 1.0 μ l, dNTP mixed solution (2.5mM each) 0.8 μ l, 10X PCR reaction buffer (Mg
2+Free) each 0.2 μ l, MgCl of 1.0 μ l, upstream primer and downstream primer (10 μ M)
2(25mM) 1.0 μ l, T aq archaeal dna polymerase (5Units/ μ l) 0.08 μ l, deionized water 5.72 μ l.Reaction conditions is 94 ℃, 12 minutes, carries out 94 ℃ of 30 round-robin, 30 seconds, 60 ℃, 30 seconds, 72 ℃, 30 seconds, carries out then 72 ℃, 10 minutes.
(3) PCR product purification:
Each reaction system is cumulative volume 12.5 μ l, comprises PCR product 10 μ l, SAP enzyme (1units/ μ 1) 0.375 μ l, ExoI enzyme (10units/ μ l) 0.1875 μ l, deionized water 1.9375 μ l.Reaction conditions is 37 ℃, 15 minutes, 72 ℃, 20 minutes.
(4) dna sequencing reaction
The system of each reaction is cumulative volume 5 μ l, comprises PCR purified product 1 μ l, BigDye mix (25%) 1 μ l, dna sequencing primer (3.2 μ M) 1 μ l, deionized water 2 μ l.Reaction conditions is 98 ℃, 2 minutes, carries out 96 ℃ of 25 round-robin, 30 seconds, 55 ℃, 30 seconds, and 60 ℃, 4 minutes.
Reaction finishes the every pipe in back and adds EDTA solution (125mM) 1 μ l and ethanolic soln (100%) 15 μ l, and precipitation is 15 minutes under room temperature; Centrifugal 30 minutes of 4 ℃ of rotating speeds, remove supernatant liquor gently with 3650 rev/mins; Add ethanolic soln (70%) 30 μ l, with 3650 rev/mins rotating speed centrifugal 15 minutes, remove supernatant liquor gently; Room temperature is placed and is added HIDI solution 8 μ l after 20 minutes, puts into sequenator.
Test kit of the present invention is used to detect the mutant of duchenne muscular dystrophy Disease-causing gene DMD, can find the carrier and the patient of Disease-causing gene quickly and easily, can be applicable to antenatal diagnosis, stop the infant birth, reduce the sickness rate of duchenne muscular dystrophy.
Embodiment
Embodiment 1
One person-portion detects the test kit whether duchenne muscular dystrophy Disease-causing gene DMD undergos mutation, and comprises following composition:
(1) DMD gene the 1st to the 79th exon district pcr amplification and sequencing primer are 79 pairs, every primer 1 OD, and each bar primer sequence is referring to table 1;
(2) pcr amplification reagent: dNTP mixed solution (2.5mM each) 72 μ l, 10X PCR reaction buffer (Mg
2+Free) 90 μ l, MgCl
2(25mM) 90 μ l, T aq archaeal dna polymerase (5Units/ μ l) 7.2 μ l;
(3) PCR product purification reagent: SAP enzyme (1units/ μ l) 33.75 μ l, ExoI enzyme (10units/ μ l) 16.875 μ l;
(4) sequencing reagent, BigDye mix (25%) 90 μ l, EDTA solution (125mM) 90 μ l, ethanolic soln (100%) 1.35ml, ethanolic soln (70%) 2.7ml, HIDI solution 720 μ l.
This test kit is stored in-20 ℃, reduces multigelation as far as possible.
Embodiment 2
The use step of the test kit whether one person-portion detection duchenne muscular dystrophy Disease-causing gene DMD undergos mutation comprises:
(1) genomic dna of extraction sample;
(2) DMD gene PCR amplification:
79 pairs of primers carry out 79 PCR reactions altogether.Each reaction system is cumulative volume 10 μ l, comprises genomic dna (100ng/ μ l) 1.0 μ l, dNTP mixed solution (2.5mM each) 0.8 μ l, 10X PCR reaction buffer (Mg
2+Free) each 0.2 μ l, MgCl of 1.0 μ l, upstream primer and downstream primer (10 μ M)
2(25mM) 1.0 μ l, T aq archaeal dna polymerase (5Units/ μ l) 0.08 μ l, deionized water 5.72 μ l.Reaction conditions is 94 ℃, 12 minutes, carries out 94 ℃ of 30 round-robin, 30 seconds, 60 ℃, 30 seconds, 72 ℃, 30 seconds, carries out then 72 ℃, 10 minutes.
(3) PCR product purification:
Carry out 79 PCR purification reactions altogether.Each reaction system is cumulative volume 12.5 μ l, comprises PCR product 10 μ l, SAP enzyme (1units/ μ l) 0.375 μ l, ExoI enzyme (10units/ μ l) 0.1875 μ l, deionized water 1.9375 μ l.Reaction conditions is 37 ℃, 15 minutes, 72 ℃, 20 minutes.
(4) dna sequencing reaction
Carry out 79 dna sequencing reactions altogether.The system of each reaction is cumulative volume 5 μ l, comprises PCR purified product 1 μ l, BigDye mix (25%) 1 μ l, dna sequencing primer (3.2 μ M) 1 μ l, deionized water 2 μ l.Reaction conditions is 98 ℃, 2 minutes, carries out 96 ℃ of 25 round-robin, 30 seconds, 55 ℃, 30 seconds, and 60 ℃, 4 minutes.Reaction finishes the back and adds EDTA solution (125mM) 1 μ l and ethanolic soln (100%) 15 μ l, and precipitation is 15 minutes under room temperature; Centrifugal 30 minutes of 4 ℃ of rotating speeds, remove supernatant liquor gently with 3650 rev/mins; Add ethanolic soln (70%) 30 μ l, with 3650 rev/mins rotating speed centrifugal 15 minutes, remove supernatant liquor gently; Room temperature is placed and is added HIDI solution 8 μ l after 20 minutes, puts into sequenator and checks order.
＜110〉Shanghai one hundred true bio tech ltd
＜120〉duchenne muscular dystrophy Disease-causing gene sudden change detection kit and application
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<211>?22
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?82
ttcagctcat?ttgtctgaat?tg 22
<210>?83
<211>?21
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?83
aaactctgca?aatgcaggaa?a 21
<210>?84
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?84
ttccatcacc?cttcagaacc 20
<210>?85
<211>?22
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?85
ttctttgcca?gtacaactgc?at 22
<210>?86
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?86
tagtgccttt?caccctgctt 20
<210>?87
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?87
aaattgccat?gtttgtgtcc 20
<210>?88
<211>?22
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?88
gaactgcagg?gttaagaaga?aa 22
<210>?89
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?89
ctcggtcaag?tcgcttcatt 20
<210>?90
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?90
atccacatac?cagcctcctc 20
<210>?91
<211>?23
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?91
tgctgctaaa?ataacacaaa?tca 23
<210>?92
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?92
agtggtccct?gtgcctattg 20
<210>?93
<211>?22
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?93
tgatctgcaa?tacatgtgga?gt 22
<210>?94
<211>?25
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?94
cactgattat?aaatagtcca?cgtca 25
<210>?95
<211>?25
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?95
tgcaaccaca?atagatactc?ttcaa 25
<210>?96
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?96
tgctcttctc?ttccttgtgg 20
<210>?97
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?97
ttgtccaggc?atgagaatga 20
<210>?98
<211>?21
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?98
acagttgcct?aagaactggt?g 21
<210>?99
<211>?22
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?99
tggctggtct?cacaattgta?ct 22
<210>?100
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?100
ggagagactg?gcatctccat 20
<210>?101
<211>?22
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?101
tcctgttgtt?catcatccta?gc 22
<210>?102
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?102
atgattgtgc?cactgcactt 20
<210>?103
<211>?21
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?103
gacctgagga?ttcagaagct?g 21
<210>?104
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?104
gtctgagcca?agtccgtgag 20
<210>?105
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?105
tgtcattctt?gcatgccttc 20
<210>?106
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?106
cctctgcctc?tctcctcctt 20
<210>?107
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?107
agaaggcaat?acgccaagaa 20
<210>?108
<211>?26
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?108
ttacgtagac?atgtgagata?ccagtt 26
<210>?109
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?109
tctcagcatt?ctgcatgtgg 20
<210>?110
<211>?24
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?110
ttgttgatgt?cttgatgagt?acga 24
<210>?111
<211>?23
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?111
ggcattaact?atccagaatg?ctc 23
<210>?112
<211>?23
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?112
ttgcaacaat?agaattcctc?ctc 23
<210>?113
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?113
ttgtggtatg?catcctaacg 20
<210>?114
<211>?24
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?114
aataccatgt?gaatgaagac?tgaa 24
<210>?115
<211>?24
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?115
tcctaatacc?attgccttag?gtgt 24
<210>?116
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?116
tacagcctgc?agaactgagc 20
<210>?117
<211>?25
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?117
aacgagtctg?gaatactata?tacgg 25
<210>?118
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?118
atccaattgg?ccttcctctt 20
<210>?119
<211>?21
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?119
cagcaggtta?atccatcctg?t 21
<210>?120
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?120
tgtaggccag?gctaatgtcg 20
<210>?121
<211>?22
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?121
tgtagtgttc?cacattggat?gg 22
<210>?122
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?122
ttggatagga?aggtgccact 20
<210>?123
<211>?22
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?123
ttgtggtggt?tgtagttatt?gg 22
<210>?124
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?124
gcaacaggaa?ttgtgacagc 20
<210>?125
<211>?24
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?125
tcatcaactt?agagtcgagg?cata 24
<210>?126
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?126
gcattaggtc?cacagcttcc 20
<210>?127
<211>?21
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?127
ccattgcatt?gtaggtcatc?a 21
<210>?128
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?128
catgaggaat?ctgctgtcaa 20
<210>?129
<211>?23
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?129
ttgagaattg?ctactggaat?tga 23
<210>?130
<211>?24
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?130
catacctact?gcctactgaa?gagc 24
<210>?131
<211>?23
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?131
aagtctcgct?caggtgtgta?gtt 23
<210>?132
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?132
taacagcaac?tggcacagga 20
<210>?133
<211>?27
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?133
ttccagtacc?ttcctactta?atctcaa 27
<210>?134
<211>?21
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?134
gcctacagtt?gagagccact?t 21
<210>?135
<211>?21
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?135
ttgctcacaa?ccacaacact?c 21
<210>?136
<211>?19
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?136
gtggcaactg?gacatcagc 19
<210>?137
<211>?23
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?137
tggttcatac?ttaattggct?gga 23
<210>?138
<211>?19
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?138
ttcacctggt?ggaggaacc 19
<210>?139
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?139
aagcattcta?ggccatgtgt 20
<210>?140
<211>?27
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?140
agttgattag?taggaatcag?acaagtt 27
<210>?141
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?141
tggttaagcc?aacacacaca 20
<210>?142
<211>?25
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?142
tgatgctaat?tcctatatcc?tgtgc 25
<210>?143
<211>?21
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?143
tggtagatca?caacctcagc?a 21
<210>?144
<211>?21
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?144
ggagacacat?gcaacaagtc?a 21
<210>?145
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?145
ttcttcggtg?gcagtcagat 20
<210>?146
<211>?21
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?146
gaattgatgc?ttgttgcacc?t 21
<210>?147
<211>?22
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?147
tctcattcca?gtgtagctct?ga 22
<210>?148
<211>?19
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?148
gccagataga?gctgccaga 19
<210>?149
<211>?19
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?149
agatcgtgcc?actgcactc 19
<210>?150
<211>?21
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?150
tgatactgcg?tgttggctt?c 21
<210>?151
<211>?22
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?151
aatgacgtct?tctggatcat?gt 22
<210>?152
<211>?19
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?152
gcgtgcacac?acacaattc 19
<210>?153
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?153
cgatgcagct?tctgtgttgt 20
<210>?154
<211>?21
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?154
caatcaatca?accaaccaac?c 21
<210>?155
<211>?23
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?155
ttccactgac?aacgaaagta?aag 23
<210>?156
<211>?23
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?156
cacggtggta?tctattgaat?gaa 23
<210>?157
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?157
tgcgagtagt?tccacacagg 20
<210>?158
<211>?20
<212>?DNA
＜213〉artificial sequence
<220>
＜223〉primer
<400>?158
tgcataactg?ggtgcagaga 20
Claims (2)
1. one kind is detected the test kit whether duchenne muscular dystrophy Disease-causing gene DMD gene undergos mutation, it is characterized in that comprising following composition: 79 pairs of (1) DMD gene the 1st to the 79th exon district pcr amplification and sequencing primers, (2) pcr amplification reagent, (3) PCR product purification reagent, (4) dna sequencing reagent;
Described 79 pairs of primer sequences are as follows:
The 1st pair: F:5 '-tggcgaggaactctactattg-3 ', R:5 '-ttgaagagctcggcaacaa-3 '
The 2nd pair: F:5 '-aagtgatccgtcctcactgg-3 ', R:5 '-ccatatcttctgctgcttactcc-3 '
The 3rd pair: F:5 '-tcggcagattaattatgcacac-3 ', R:5 '-tctgcaggcggtagagtatg-3 '
The 4th pair: F:5 '-atgcctcacaggctctgttc-3 ', R:5 '-ccacaatcaggcatacacga-3 '
The 5th pair: F:5 '-aagcttcaatgctaagtctctga-3 ', R:5 '-cagacgacatggtagtgtcaat-3 '
The 6th pair: F:5 '-tggcatagataccaatgaatcag-3 ', R:5 '-ggacatgatctggaaccatactg-3 '
The 7th pair: F:5 '-ttacactcaagacttaaggactatgg-3 ', R:5 '-agtgcaagaaggctgaacatt-3 '
The 8th pair: F:5 '-gacattcatggacaattcactg-3 ', R:5 '-atgaatattgtagtcaatgaagcaa-3 '
The 9th pair: F:5 '-tgatattaagtgccttcattctgg-3 ', R:5 '-ccagctcttcacgaggagataa-3 '
The 10th pair: F:5 '-tgccagtggacagtcctagc-3 ', R:5 '-aatgccttcaatggttgctc-3 '
The 11st pair: F:5 '-gcctgcttctgaagaacttg-3 ', R:5 '-tgagaatcgtaacttaaccatcaa-3 '
The 12nd pair: F:5 '-ccttctcaatgtccaatagatgc-3 ', R:5 '-ccatcaaccatgtcatctgtg-3 '
The 13rd pair: F:5 '-ggagtacctgagatgtagcagaa-3 ', R:5 '-cacagcacttcagctgattatga-3 '
The 14th pair: F:5 '-ccttccaacttctagcgtaca-3 ', R:5 '-ttgaataatctatgatccaagcaa-3 '
The 15th pair: F:5 '-ccaagatccgtgccttctaa-3 ', R:5 '-tcaccaccattctccaacaa-3 '
The 16th pair: F:5 '-aaggtcaatctaccaacaagcaa-3 ', R:5 '-tctcactacattaactgacattacagg-3 '
The 17th pair: F:5 '-agaggtgtcaggcaggagtc-3 ', R:5 '-cctagtcagtcacagaagaatcca-3 '
The 18th pair: F:5 '-cacgtgataagctgacagagtga-3 ', R:5 '-ttaacaagtgcttgtctgatataattc-3 '
The 19th pair: F:5 '-ttgttgtgacgcaagtctga-3 ', R:5 '-cacaatcaattagcgctgtg-3 '
The 20th pair: F:5 '-ggctggtgatagaggcttgt-3 ', R:5 '-ttcgttccttgttaatacaggttg-3 '
The 21st pair: F:5 '-ggcatattacacatgtagcagga-3 ', R:5 '-ttgagaatgtgacaggagttacaa-3 '
The 22nd pair: F:5 '-tcatctactttgtttacatgtttgaa-3 ', R:5 '-taagcaaatcgccatccttt-3 '
The 23rd pair: F:5 '-gacttcacacaccagtgctca-3 ', R:5 '-caccagtagcatctaaccacca-3 '
The 24th pair: F:5 '-atccatatgcaatgccatca-3 ', R:5 '-aggaacaaagccttaaccaaa-3 '
The 25th pair: F:5 '-tcacacttatgggaaattgagtg-3 ', R:5 '-gaaccaggaaagagcagactgt-3 '
The 26th pair: F:5 '-ctgggatgttgtgagaaagaaa-3 ', R:5 '-gaatgaggaattcaagccaaa-3 '
The 27th pair: F:5 '-acctgctacaaagtaaaggtgaa-3 ', R:5 '-aagtaccagtgctgagtgataacat-3 '
The 28th pair: F:5 '-ccgcaacactgtcataatcc-3 ', R:5 '-tggcattggattgtctctga-3 '
The 29th pair: F:5 '-ggaagctgcgaaatctgtct-3 ', R:5 '-cctccaaatagtcaaatcagtgaa-3 '
The 30th pair: F:5 '-ggtggttgaggagagtttctg-3 ', R:5 '-ctcaaatccaatcttgccaat-3 '
The 31st pair: F:5 '-tcctgtgttggatgaatgga-3 ', R:5 '-gccacaatacatgtgccaat-3 '
The 32nd pair: F:5 '-tcaaacatggaatagcaattaagg-3 ', R:5 '-gagagagagaggtgtttagaattgc-3 '
The 33rd pair: F:5 '-ggacaaacgatgtcatctgc-3 ', R:5 '-tcctgctaaagcttaccattg-3 '
The 34th pair: F:5 '-aaccttgcagaatcatagtaacca-3 ', R:5 '-gggaaagataaaggaaggaagaa-3 '
The 35th pair: F:5 '-tcttctcgctcacttgctca-3 ', R:5 '-gggaggagtggcgtttatct-3 '
The 36th pair: F:5 '-ttagcaacaggaggttgaataaa-3 ', R:5 '-ccataccatagtaattcagcatttg-3 '
The 37th pair: F:5 '-gaaaggctatgagcacagtatctta-3 ', R:5 '-ttctgatgactaagtctgaagcagat-3 '
The 38th pair: F:5 '-aaagatgagggacgcaaatta-3 ', R:5 '-caaggaaatgcatcaaatcaaa-3 '
The 39th pair: F:5 '-aagggacaacagcctttgaa-3 ', R:5 '-aagcccaaagtgagggaaac-3 '
The 40th pair: F:5 '-ttcagcccagtttgtagattca-3 ', R:5 '-agatttctgaagccaaccaca-3 '
The 41st pair: F:5 '-ttgcaacaccatttgctacc-3 ', R:5 '-ttcagctcatttgtctgaattg-3 '
The 42nd pair: F:5 '-aaactctgcaaatgcaggaaa-3 ', R:5 '-ttccatcacccttcagaacc-3 '
The 43rd pair: F:5 '-ttctttgccagtacaactgcat-3 ', R:5 '-tagtgcctttcaccctgctt-3 '
The 44th pair: F:5 '-aaattgccatgtttgtgtcc-3 ', R:5 '-gaactgcagggttaagaagaaa-3 '
The 45th pair: F:5 '-ctcggtcaagtcgcttcatt-3 ', R:5 '-atccacataccagcctcctc-3 '
The 46th pair: F:5 '-tgctgctaaaataacacaaatca-3 ', R:5 '-agtggtccctgtgcctattg-3 '
The 47th pair: F:5 '-tgatctgcaatacatgtggagt-3 ', R:5 '-cactgattataaatagtccacgtca-3 '
The 48th pair: F:5 '-tgcaaccacaatagatactcttcaa-3 ', R:5 '-tgctcttctcttccttgtgg-3 '
The 49th pair: F:5 '-ttgtccaggcatgagaatga-3 ', R:5 '-acagttgcctaagaactggtg-3 '
The 50th pair: F:5 '-tggctggtctcacaattgtact-3 ', R:5 '-ggagagactggcatctccat-3 '
The 51st pair: F:5 '-tcctgttgttcatcatcctagc-3 ', R:5 '-atgattgtgccactgcactt-3 '
The 52nd pair: F:5 '-gacctgaggattcagaagctg-3 ', R:5 '-gtctgagccaagtccgtgag-3 '
The 53rd pair: F:5 '-tgtcattcttgcatgccttc-3 ', R:5 '-cctctgcctctctcctcctt-3 '
The 54th pair: F:5 '-agaaggcaatacgccaagaa-3 ', R:5 '-ttacgtagacatgtgagataccagtt-3 '
The 55th pair: F:5 '-tctcagcattctgcatgtgg-3 ', R:5 '-ttgttgatgtcttgatgagtacga-3 '
The 56th pair: F:5 '-ggcattaactatccagaatgctc-3 ', R:5 '-ttgcaacaatagaattcctcctc-3 '
The 57th pair: F:5 '-ttgtggtatgcatcctaacg-3 ', R:5 '-aataccatgtgaatgaagactgaa-3 '
The 58th pair: F:5 '-tcctaataccattgccttaggtgt-3 ', R:5 '-tacagcctgcagaactgagc-3 '
The 59th pair: F:5 '-aacgagtctggaatactatatacgg-3 ', R:5 '-atccaattggccttcctctt-3 '
The 60th pair: F:5 '-cagcaggttaatccatcctgt-3 ', R:5 '-tgtaggccaggctaatgtcg-3 '
The 61st pair: F:5 '-tgtagtgttccacattggatgg-3 ', R:5 '-ttggataggaaggtgccact-3 '
The 62nd pair: F:5 '-ttgtggtggttgtagttattgg-3 ', R:5 '-gcaacaggaattgtgacagc-3 '
The 63rd pair: F:5 '-tcatcaacttagagtcgaggcata-3 ', R:5 '-gcattaggtccacagcttcc-3 '
The 64th pair: F:5 '-ccattgcattgtaggtcatca-3 ', R:5 '-catgaggaatctgctgtcaa-3 '
The 65th pair: F:5 '-ttgagaattgctactggaattga-3 ', R:5 '-catacctactgcctactgaagagc-3 '
The 66th pair: F:5 '-aagtctcgctcaggtgtgtagtt-3 ', R:5 '-taacagcaactggcacagga-3 '
The 67th pair: F:5 '-ttccagtaccttcctacttaatctcaa-3 ', R:5 '-gcctacagttgagagccactt-3 '
The 68th pair: F:5 '-ttgctcacaaccacaacactc-3 ', R:5 '-gtggcaactggacatcagc-3 '
The 69th pair: F:5 '-tggttcatacttaattggctgga-3 ', R:5 '-ttcacctggtggaggaacc-3 '
The 70th pair: F:5 '-aagcattctaggccatgtgt-3 ', R:5 '-agttgattagtaggaatcagacaagtt-3 '
The 71st pair: F:5 '-tggttaagccaacacacaca-3 ', R:5 '-tgatgctaattcctatatcctgtgc-3 '
The 72nd pair: F:5 '-tggtagatcacaacctcagca-3 ', R:5 '-ggagacacatgcaacaagtca-3 '
The 73rd pair: F:5 '-ttcttcggtggcagtcagat-3 ', R:5 '-gaattgatgcttgttgcacct-3 '
The 74th pair: F:5 '-tctcattccagtgtagctctga-3 ', R:5 '-gccagatagagctgccaga-3 '
The 75th pair: F:5 '-agatcgtgccactgcactc-3 ', R:5 '-tgatactgcgtgttggcttc-3 '
The 76th pair: F:5 '-aatgacgtcttctggatcatgt-3 ', R:5 '-gcgtgcacacacacaattc-3 '
The 77th pair: F:5 '-cgatgcagcttctgtgttgt-3 ', R:5 '-caatcaatcaaccaaccaacc-3 '
The 78th pair: F:5 '-ttccactgacaacgaaagtaaag-3 ', R:5 '-cacggtggtatctattgaatgaa-3 '
The 79th pair: F:5 '-tgcgagtagttccacacagg-3 ', R:5 '-tgcataactgggtgcagaga-3 '.
2. be used to detect the DMD gene mutation body according to right 1 described test kit.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011101486070A CN102230001A (en) | 2011-06-03 | 2011-06-03 | Kit for detecting mutation of duchenne muscular dystrophy causing gene and use thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011101486070A CN102230001A (en) | 2011-06-03 | 2011-06-03 | Kit for detecting mutation of duchenne muscular dystrophy causing gene and use thereof |
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| CN102230001A true CN102230001A (en) | 2011-11-02 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN2011101486070A Pending CN102230001A (en) | 2011-06-03 | 2011-06-03 | Kit for detecting mutation of duchenne muscular dystrophy causing gene and use thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104498614A (en) * | 2014-12-31 | 2015-04-08 | 广州和泰科技有限公司 | Probe and non-invasive kit for detecting pseudo-hypertrophic muscular dystrophy |
| CN105441580A (en) * | 2016-01-26 | 2016-03-30 | 绍兴华因生物科技有限公司 | Method and primers for detecting heterozygosity DMD gene deletion |
| CN108220418A (en) * | 2017-12-29 | 2018-06-29 | 东莞博奥木华基因科技有限公司 | The detection kit and method of Du Shi based on multiplex PCR capture technique/bayesian muscular dystrophy |
| CN109486936A (en) * | 2018-11-06 | 2019-03-19 | 南方医科大学南方医院 | Detect the genetic fragment of DMD deletion mutant |
| CN111139297A (en) * | 2020-04-02 | 2020-05-12 | 北京大学第三医院（北京大学第三临床医学院） | Kit for preimplantation embryo genetic diagnosis and prenatal diagnosis of DMD |
| CN113322312A (en) * | 2021-07-18 | 2021-08-31 | 华中科技大学同济医学院附属协和医院 | Detection kit and detection method for X-linked DMD gene mutant and application of detection kit and detection method |
| CN115851744A (en) * | 2022-09-21 | 2023-03-28 | 湖南家辉生物技术有限公司 | DMD gene mutant and application thereof, and primer combination, reagent and kit for detecting DMD gene mutant |
-
2011
- 2011-06-03 CN CN2011101486070A patent/CN102230001A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104498614A (en) * | 2014-12-31 | 2015-04-08 | 广州和泰科技有限公司 | Probe and non-invasive kit for detecting pseudo-hypertrophic muscular dystrophy |
| CN105441580A (en) * | 2016-01-26 | 2016-03-30 | 绍兴华因生物科技有限公司 | Method and primers for detecting heterozygosity DMD gene deletion |
| CN105441580B (en) * | 2016-01-26 | 2018-10-16 | 绍兴华因生物科技有限公司 | Detect the method and the primer of heterozygosity DMD gene delections |
| CN108220418A (en) * | 2017-12-29 | 2018-06-29 | 东莞博奥木华基因科技有限公司 | The detection kit and method of Du Shi based on multiplex PCR capture technique/bayesian muscular dystrophy |
| CN108220418B (en) * | 2017-12-29 | 2018-11-09 | 东莞博奥木华基因科技有限公司 | The detection kit and method of Du Shi based on multiplex PCR capture technique/bayesian muscular dystrophy |
| CN109486936A (en) * | 2018-11-06 | 2019-03-19 | 南方医科大学南方医院 | Detect the genetic fragment of DMD deletion mutant |
| CN111139297A (en) * | 2020-04-02 | 2020-05-12 | 北京大学第三医院（北京大学第三临床医学院） | Kit for preimplantation embryo genetic diagnosis and prenatal diagnosis of DMD |
| CN113322312A (en) * | 2021-07-18 | 2021-08-31 | 华中科技大学同济医学院附属协和医院 | Detection kit and detection method for X-linked DMD gene mutant and application of detection kit and detection method |
| CN115851744A (en) * | 2022-09-21 | 2023-03-28 | 湖南家辉生物技术有限公司 | DMD gene mutant and application thereof, and primer combination, reagent and kit for detecting DMD gene mutant |
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Application publication date: 20111102 |