具体实施方式
本发明涉及一个基于二代测序技术检测微单倍型基因座的引物组合物,其包括163个微单倍型基因座的扩增引物一对或多对;
其中,所述的163个微单倍型基因座均源自ALFRED网站中已收录的微单倍型基因座和文献中已发表的基因座,分布于内含子区域、在中国人群中多态性较好、分布长度≤300bp,其名称、染色体信息及其位点信息如表1所示:
表1 163个微单倍型基因座的名称、染色体信息及包含的SNP信息
根据筛选微单倍型基因座的物理位置,采用在线引物设计工具Ion AmpliSeqDesigner设计多重PCR引物。设计原则包括:(1)最优的熔链温度;(2)避免引物二聚体和发夹结构;(3)GC含量在20%~80%之间;(4)进行脱靶分析以减少引物脱靶杂交;(5)进行重叠分析以减少引物的数量。在本发明一优选的实施方式中,上述引物组合物包括核苷酸序列为SEQ ID No.1~326的引物一对或多对,具体的引物序列信息如下:
表2 163个微单倍型基因座的扩增引物序列号以及引物序列
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在本发明一优选的实施方式中,上述引物组合物包括核苷酸序列为SEQ ID No.1~326的引物。
本发明还涉及一种基于二代测序技术检测微单倍型基因座的试剂盒,包括上述引物组合物,其还包括PCR混合液和PCR反应液。
下面通过具体实施例和附图对本发明进行详细和具体的介绍,以使更好的理解本发明,但是下述实施例并不限制本发明范围。
实施例中方法如无特殊说明的采用常规方法,使用的试剂如无特殊说明的使用常规市售试剂或按常规方法配制的试剂。
实施例1
本实施例提供一种利用上述引物组合物或者试剂盒的基于二代测序技术检测微单倍型基因座的方法,包括如下步骤:
(1)取待测样品,提取样本DNA,定量样本浓度为5ng/μL;
(2)进行第一轮多重PCR,PCR扩增体系及扩增条件如表3所示。
表3文库构建第一轮PCR复合扩增反应体系
PCR反应条件为:95℃,预变性15分钟;95℃,变性30秒,60℃,退火90秒,72℃,延伸30s,循环24次;72℃,保温10分钟。反应完成后,向其中加入1μL纯化反应液纯化产物,并完成如下反应:37℃,10分钟,50℃,10分钟,65℃,10分钟,4℃,保温。然后进行磁珠分选。
(3)补平修复加A:反应体系如表4所示:
表4文库构建补平修复加A反应体系
PCR反应条件为:30℃,30分钟,65℃,30分钟,4℃,保温。
(4)接头连接:反应体系如表5所示:
表5文库构建接头连接反应体系
PCR反应条件为:25℃,15分钟,4℃,保温。然后利用纯化磁珠纯化反应产物。
(5)再次进行PCR扩增,PCR反应体系如表6所示:
表6文库构建第二轮PCR反应体系
PCR反应条件为:37℃,15分钟;98℃,预变性45秒;98℃,变性15秒,60℃,退火30秒,72℃,延伸30秒,循环10次;72℃,5分钟;4℃,保温。
(6)文库的纯化和定量:再次利用纯化磁珠纯化产物,采用Qubit进行文库定量及质控。
(7)上机测序及数据分析:将构建好的文库放置于MiSeq FGxTM平台进行测序分析;对于获得的测序数据,采用Trimmomatic软件去测序接头,然后采用BWA软件进行序列比对将序列与人类参考基因组(hg19)进行比对,利用Python工具获取微单倍型。
该方法可用于个人识别和亲缘鉴定,具体是挑选多态性较好的48个微单倍型基因座:MH01CP008、MH01CP012、MH01CP016、MH01KK117、MH01KK205、MH01KK211、MH02KK134、MH02KK136、MH04CP002、MH04CP003、MH04CP007、MH04KK030、MH05CP004、MH05CP006、MH05KK020、MH05KK170、MH06CP003、MH06CP007、MH09KK153、MH10CP003、MH10KK163、MH11CP003、MH11CP005、MH11KK180、MH12KK046、MH12KK202、MH13CP008、MH13KK213、MH13KK217、MH13KK218、MH13KK225、MH14CP003、MH14CP004、MH15CP001、MH15KK066、MH16KK255、MH16KK302、MH17CP001、MH17CP006、MH17KK272、MH18CP003、MH18CP005、MH19CP007、MH19KK299、MH20KK058、MH20KK307、MH21KK315、MH21KK324,采用对应表2中的引物序列按照上述步骤进行检测分析。
该方法可用于族源推断,具体是基于全部163个基因座的微单倍型分型结果进行分析。
实施例2
本实施例为实施例1提供的方法的法医学验证工作,具体的实验和结果如下:
按照DNA分析方法科学工作组(Scientific Working Group for DNA AnalysisMethods,SWGDAM)要求对实施例1构建的复合扩增体系进行灵敏度、准确性、重复性以及法医学参数计算。
结果显示实施例1构建的方法(针对163个微单倍型基因座)具有高灵敏度,所有浓度均可以获得微单倍型基因座的完整基因分型,不同浓度梯度DNA投入量下二代测序数据统计见图1和图2;准确性高,采用Sanger测序法进行验证,结果显示微单倍型体系中所有SNP位点分型均与二代测序结果一致;重复性好。
针对48个多态性好的微单倍型基因座,48个基因座平均杂合度达到0.7227,多态性信息含量均大于0.60,平均个体识别概率达到0.8692,累计个人识别概率为1-8.26×10-44,二联体累积非父排除率和三联体累积非父排除率分别为1~1.26×10-8和1~8.27×10-16。
实施例3
本实施例为实施例1提供的方法与二代测序平台的STR试剂盒对混合物样本分析效能的比较。
基于二代测序平台的ForenSeqTM DNA Signature Prep Kit中常染色体STR基因座,由于其高敏感性以及stutter峰扩增的影响,在20:1的混合比以下开始出现次要贡献者等位基因的大量丢失。制备不同混合比例的DNA混合物样本,分别使用实施例1提供的方法(针对163个微单倍型基因座)与ForenSeqTM DNA Signature Prep Kit进行检测,比较两者的混合物检测效能。表7是两者在不同混合比例的DNA混合物样本中的次要贡献者独特等位基因检出率。结果显示,实施例1提供的方法对混合物的检测效果明显优于二代测序平台的STR试剂盒。
表7两种遗传标记在不同混合比例的DNA混合物样本中的次要贡献者独特等位基因检出率
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实施例4
本实施例为实施例1提供的方法在族源推断方面的应用,具体的操作步骤和结果如下:
利用千人基因组计划中26个群体联合华东汉族群体共27个群体的微单倍型分型数据,比较27个群体间基因型频率分布差异,计算微单倍型基因座在27个群体中的In值,评估基因座的族源信息含量,并进行主成分分析。
结果显示,163个微单倍型基因座的In值均大于0.185,具备较高的祖先信息含量,可用于族源推断。由图3可知,本发明所包含的163个微单倍型基因座可将世界范围主要地区的人群较为清晰地区分,非洲、东亚、南亚与欧洲群体之间有明显的分离。
由上述实施例可知,本发明提供的引物组合物、试剂盒和方法为我国法医学研究中个体识别、亲子鉴定、混合物分析以及族源推断等法医学实际工作提供了新的检测手段。
以上对本发明的具体实施例进行了详细描述,但其只作为范例,本发明并不限制于以上描述的具体实施例。对于本领域技术人员而言,任何对本发明进行等同修改和替代也都在本发明的范畴之中。因此,在不脱离本发明精神和范围下所作的均等变换和修改,都应涵盖在本发明的范围内。
序列表
<110> 司法鉴定科学研究院
<120> 一种基于二代测序技术检测微单倍型基因座的引物组合物、试剂盒和方法及其应用
<160> 326
<170> SIPOSequenceListing 1.0
<210> 1
<211> 22
<212> DNA
<213> MH01CP007的扩增引物(Artificial Sequence)
<400> 1
ttctccccaa atcacagcac cc 22
<210> 2
<211> 23
<212> DNA
<213> MH01CP007的扩增引物(Artificial Sequence)
<400> 2
cgtaaggatg ggcaaaacgt tca 23
<210> 3
<211> 28
<212> DNA
<213> MH01CP008的扩增引物(Artificial Sequence)
<400> 3
aagcagtttg atgtgagctc taaaacga 28
<210> 4
<211> 28
<212> DNA
<213> MH01CP008的扩增引物(Artificial Sequence)
<400> 4
gccagtagaa attctaaaac aaaaccca 28
<210> 5
<211> 23
<212> DNA
<213> MH01CP012的扩增引物(Artificial Sequence)
<400> 5
atcattttct cagtgcgcaa cac 23
<210> 6
<211> 28
<212> DNA
<213> MH01CP012的扩增引物(Artificial Sequence)
<400> 6
ctttgatgtc agattttctt aggaccga 28
<210> 7
<211> 24
<212> DNA
<213> MH01CP016的扩增引物(Artificial Sequence)
<400> 7
cactcacttt gtgaccattc cggt 24
<210> 8
<211> 26
<212> DNA
<213> MH01CP016的扩增引物(Artificial Sequence)
<400> 8
ctgaaggact actacctctt ctacct 26
<210> 9
<211> 22
<212> DNA
<213> MH01KK001的扩增引物(Artificial Sequence)
<400> 9
gatgagcacc tcgagaagac ct 22
<210> 10
<211> 22
<212> DNA
<213> MH01KK001的扩增引物(Artificial Sequence)
<400> 10
gatggctggt accgatcatc tc 22
<210> 11
<211> 24
<212> DNA
<213> MH01KK070的扩增引物(Artificial Sequence)
<400> 11
tagcaacgcc aatctcagag aggt 24
<210> 12
<211> 28
<212> DNA
<213> MH01KK070的扩增引物(Artificial Sequence)
<400> 12
tgctgtaagc actctacaca tatcaatt 28
<210> 13
<211> 25
<212> DNA
<213> MH01KK072的扩增引物(Artificial Sequence)
<400> 13
ataagctatg ctgagggaag tctgg 25
<210> 14
<211> 22
<212> DNA
<213> MH01KK072的扩增引物(Artificial Sequence)
<400> 14
atgaagctgg ctcagtcaac tc 22
<210> 15
<211> 26
<212> DNA
<213> MH01KK106的扩增引物(Artificial Sequence)
<400> 15
catagtttcc agagtggttt gcaggc 26
<210> 16
<211> 23
<212> DNA
<213> MH01KK106的扩增引物(Artificial Sequence)
<400> 16
atgagatggg tggtggacag tta 23
<210> 17
<211> 23
<212> DNA
<213> MH01KK117的扩增引物(Artificial Sequence)
<400> 17
tcctaggcgt aaatggatga gag 23
<210> 18
<211> 26
<212> DNA
<213> MH01KK117的扩增引物(Artificial Sequence)
<400> 18
atgatagaat gtagaaccca gccatc 26
<210> 19
<211> 26
<212> DNA
<213> MH01KK172的扩增引物(Artificial Sequence)
<400> 19
cttaatgata atgctggcag agtctg 26
<210> 20
<211> 28
<212> DNA
<213> MH01KK172的扩增引物(Artificial Sequence)
<400> 20
ttgatatatt tccaaacacc tgtgtgct 28
<210> 21
<211> 25
<212> DNA
<213> MH01KK205的扩增引物(Artificial Sequence)
<400> 21
atctttaaga gtccgctttg tgttt 25
<210> 22
<211> 25
<212> DNA
<213> MH01KK205的扩增引物(Artificial Sequence)
<400> 22
aatgtctccc tgaggaattc tacct 25
<210> 23
<211> 28
<212> DNA
<213> MH01KK210的扩增引物(Artificial Sequence)
<400> 23
gcaagatacc aagttcttga ataaggag 28
<210> 24
<211> 24
<212> DNA
<213> MH01KK210的扩增引物(Artificial Sequence)
<400> 24
cacctcctcc ataatccaca agtg 24
<210> 25
<211> 26
<212> DNA
<213> MH01KK211的扩增引物(Artificial Sequence)
<400> 25
cacaaaatga gaggaaggtt actgag 26
<210> 26
<211> 24
<212> DNA
<213> MH01KK211的扩增引物(Artificial Sequence)
<400> 26
caaaggaggt cacatcacca tctc 24
<210> 27
<211> 28
<212> DNA
<213> MH02CP004的扩增引物(Artificial Sequence)
<400> 27
gaatctactt cacttgaatg catgttaa 28
<210> 28
<211> 27
<212> DNA
<213> MH02CP004的扩增引物(Artificial Sequence)
<400> 28
ggagaaacta agccatatat ccatggt 27
<210> 29
<211> 27
<212> DNA
<213> MH02KK003的扩增引物(Artificial Sequence)
<400> 29
tcaatcacca tgttttgact cagttta 27
<210> 30
<211> 27
<212> DNA
<213> MH02KK003的扩增引物(Artificial Sequence)
<400> 30
aattccctca gagagattat tcgatgc 27
<210> 31
<211> 27
<212> DNA
<213> MH02KK004的扩增引物(Artificial Sequence)
<400> 31
gattgttcta tgatgctggg taggggg 27
<210> 32
<211> 25
<212> DNA
<213> MH02KK004的扩增引物(Artificial Sequence)
<400> 32
tgtgttcagg ataccatgcc attag 25
<210> 33
<211> 23
<212> DNA
<213> MH02KK073的扩增引物(Artificial Sequence)
<400> 33
aggaaggcta atgacctcgc aat 23
<210> 34
<211> 27
<212> DNA
<213> MH02KK073的扩增引物(Artificial Sequence)
<400> 34
gacaccacca gaacttcttg cttatta 27
<210> 35
<211> 27
<212> DNA
<213> MH02KK102的扩增引物(Artificial Sequence)
<400> 35
tctcacttat gatgctgcta gactgac 27
<210> 36
<211> 23
<212> DNA
<213> MH02KK102的扩增引物(Artificial Sequence)
<400> 36
aagagcacat gagatccgca atc 23
<210> 37
<211> 24
<212> DNA
<213> MH02KK105的扩增引物(Artificial Sequence)
<400> 37
ggagcttgct agagaagatc acgg 24
<210> 38
<211> 25
<212> DNA
<213> MH02KK105的扩增引物(Artificial Sequence)
<400> 38
attgctcagc cacaaaagat tctca 25
<210> 39
<211> 29
<212> DNA
<213> MH02KK131的扩增引物(Artificial Sequence)
<400> 39
tttaatagtg aaagcagcaa ggttcttca 29
<210> 40
<211> 28
<212> DNA
<213> MH02KK131的扩增引物(Artificial Sequence)
<400> 40
ttttcccaga taaatttcag tgtcagct 28
<210> 41
<211> 21
<212> DNA
<213> MH02KK134的扩增引物(Artificial Sequence)
<400> 41
aaagagttgc atgccgtctg t 21
<210> 42
<211> 27
<212> DNA
<213> MH02KK134的扩增引物(Artificial Sequence)
<400> 42
gttctaggtg tcgtttgcct taagtta 27
<210> 43
<211> 27
<212> DNA
<213> MH02KK136的扩增引物(Artificial Sequence)
<400> 43
agttctcaaa gacttcaaga caagtta 27
<210> 44
<211> 28
<212> DNA
<213> MH02KK136的扩增引物(Artificial Sequence)
<400> 44
tcttttctcc acttttcaga cttcttgt 28
<210> 45
<211> 28
<212> DNA
<213> MH02KK138的扩增引物(Artificial Sequence)
<400> 45
accatctcag tgctgaaaga aatataaa 28
<210> 46
<211> 25
<212> DNA
<213> MH02KK138的扩增引物(Artificial Sequence)
<400> 46
ccagactcat cacgtcatcc agata 25
<210> 47
<211> 25
<212> DNA
<213> MH02KK139的扩增引物(Artificial Sequence)
<400> 47
gtgacagcta ggtttcatta ctgcg 25
<210> 48
<211> 26
<212> DNA
<213> MH02KK139的扩增引物(Artificial Sequence)
<400> 48
aagccaggat ttacccattt atggag 26
<210> 49
<211> 25
<212> DNA
<213> MH02KK201的扩增引物(Artificial Sequence)
<400> 49
ccaagctccc tgtgatattt ctaaa 25
<210> 50
<211> 27
<212> DNA
<213> MH02KK201的扩增引物(Artificial Sequence)
<400> 50
actggaagag tcttttgttt catagcc 27
<210> 51
<211> 26
<212> DNA
<213> MH02KK202的扩增引物(Artificial Sequence)
<400> 51
gccttttccc cttattcttt aaacaa 26
<210> 52
<211> 28
<212> DNA
<213> MH02KK202的扩增引物(Artificial Sequence)
<400> 52
tgttatctca ccactcacac attaactt 28
<210> 53
<211> 23
<212> DNA
<213> MH02KK213的扩增引物(Artificial Sequence)
<400> 53
ctcagtagtg aactgcctca ggg 23
<210> 54
<211> 28
<212> DNA
<213> MH02KK213的扩增引物(Artificial Sequence)
<400> 54
ccttccccaa cactctctaa atatttgc 28
<210> 55
<211> 25
<212> DNA
<213> MH02KK215的扩增引物(Artificial Sequence)
<400> 55
atgcaacact gcacctgaga atatg 25
<210> 56
<211> 26
<212> DNA
<213> MH02KK215的扩增引物(Artificial Sequence)
<400> 56
taccccctaa aaggttttga atgcag 26
<210> 57
<211> 26
<212> DNA
<213> MH03KK006的扩增引物(Artificial Sequence)
<400> 57
aaccaactaa tctactgaag gactgg 26
<210> 58
<211> 24
<212> DNA
<213> MH03KK006的扩增引物(Artificial Sequence)
<400> 58
caagagggac accatatgtc aagg 24
<210> 59
<211> 26
<212> DNA
<213> MH03KK007的扩增引物(Artificial Sequence)
<400> 59
catttttgaa ggctcccata ttgcat 26
<210> 60
<211> 26
<212> DNA
<213> MH03KK007的扩增引物(Artificial Sequence)
<400> 60
aaatgtgcag aaagattcca aaggag 26
<210> 61
<211> 23
<212> DNA
<213> MH03KK008的扩增引物(Artificial Sequence)
<400> 61
aggtacccat caacctcttt gtt 23
<210> 62
<211> 25
<212> DNA
<213> MH03KK008的扩增引物(Artificial Sequence)
<400> 62
acctatgtgg ctgtacaatt tgtcc 25
<210> 63
<211> 26
<212> DNA
<213> MH03KK009的扩增引物(Artificial Sequence)
<400> 63
gaagtctact cacctccagg tatacc 26
<210> 64
<211> 26
<212> DNA
<213> MH03KK009的扩增引物(Artificial Sequence)
<400> 64
ccaagcccca aagagtctga ttttat 26
<210> 65
<211> 26
<212> DNA
<213> MH03KK216的扩增引物(Artificial Sequence)
<400> 65
aagagctgaa acaagagcat tgtgca 26
<210> 66
<211> 27
<212> DNA
<213> MH03KK216的扩增引物(Artificial Sequence)
<400> 66
ccacattgta actcctagac caagaag 27
<210> 67
<211> 27
<212> DNA
<213> MH04CP002的扩增引物(Artificial Sequence)
<400> 67
acacagagtt taaggttcct tccagaa 27
<210> 68
<211> 26
<212> DNA
<213> MH04CP002的扩增引物(Artificial Sequence)
<400> 68
gggtcacttc aggataataa gctcct 26
<210> 69
<211> 26
<212> DNA
<213> MH04CP003的扩增引物(Artificial Sequence)
<400> 69
gatttgtgtc ttctgcattc acagct 26
<210> 70
<211> 22
<212> DNA
<213> MH04CP003的扩增引物(Artificial Sequence)
<400> 70
ggctgctctt gtacagcatc tc 22
<210> 71
<211> 26
<212> DNA
<213> MH04CP007的扩增引物(Artificial Sequence)
<400> 71
taaatactgt ctgcccatga ctcctc 26
<210> 72
<211> 27
<212> DNA
<213> MH04CP007的扩增引物(Artificial Sequence)
<400> 72
agagctttgg ttttaatgct attccct 27
<210> 73
<211> 28
<212> DNA
<213> MH04KK010的扩增引物(Artificial Sequence)
<400> 73
tcactatatt tttgaggaca ccaaccac 28
<210> 74
<211> 27
<212> DNA
<213> MH04KK010的扩增引物(Artificial Sequence)
<400> 74
tgttggtgcc aagtacatct ataagaa 27
<210> 75
<211> 33
<212> DNA
<213> MH04KK011的扩增引物(Artificial Sequence)
<400> 75
ttttaagaaa gaataaagaa ggacagaaag cca 33
<210> 76
<211> 32
<212> DNA
<213> MH04KK011的扩增引物(Artificial Sequence)
<400> 76
gatcatgcta tcactaagaa aattatggca aa 32
<210> 77
<211> 23
<212> DNA
<213> MH04KK013的扩增引物(Artificial Sequence)
<400> 77
tgtctaatgg ccgctgtagt aaa 23
<210> 78
<211> 27
<212> DNA
<213> MH04KK013的扩增引物(Artificial Sequence)
<400> 78
cttggcaatt taagatgctc aggaatt 27
<210> 79
<211> 28
<212> DNA
<213> MH04KK015的扩增引物(Artificial Sequence)
<400> 79
aattctatct catccatctt gagtgcat 28
<210> 80
<211> 25
<212> DNA
<213> MH04KK015的扩增引物(Artificial Sequence)
<400> 80
tattacagag tgctgcaggt cattc 25
<210> 81
<211> 28
<212> DNA
<213> MH04KK016的扩增引物(Artificial Sequence)
<400> 81
caaagctagt ttctaagtaa gccattgc 28
<210> 82
<211> 28
<212> DNA
<213> MH04KK016的扩增引物(Artificial Sequence)
<400> 82
tttttgccag agtttttagt gtactcct 28
<210> 83
<211> 28
<212> DNA
<213> MH04KK017的扩增引物(Artificial Sequence)
<400> 83
ataatggttg aagggtagaa tacacgca 28
<210> 84
<211> 24
<212> DNA
<213> MH04KK017的扩增引物(Artificial Sequence)
<400> 84
tcgttcagat gagcatgtgg ttag 24
<210> 85
<211> 23
<212> DNA
<213> MH04KK019的扩增引物(Artificial Sequence)
<400> 85
tacttgtagc agagggcctt atc 23
<210> 86
<211> 26
<212> DNA
<213> MH04KK019的扩增引物(Artificial Sequence)
<400> 86
gttagacaga agttaggcat ggagtt 26
<210> 87
<211> 28
<212> DNA
<213> MH04KK028的扩增引物(Artificial Sequence)
<400> 87
taatggaagt actgtttcag ttctgcaa 28
<210> 88
<211> 28
<212> DNA
<213> MH04KK028的扩增引物(Artificial Sequence)
<400> 88
aaaaatgttt tccttttctt cctagggc 28
<210> 89
<211> 26
<212> DNA
<213> MH04KK029的扩增引物(Artificial Sequence)
<400> 89
catttaccaa tgttggctaa tacaca 26
<210> 90
<211> 24
<212> DNA
<213> MH04KK029的扩增引物(Artificial Sequence)
<400> 90
agaacagcat aggaaggcac ttag 24
<210> 91
<211> 27
<212> DNA
<213> MH04KK030的扩增引物(Artificial Sequence)
<400> 91
aaattttggg tcttaccatg gtttcaa 27
<210> 92
<211> 24
<212> DNA
<213> MH04KK030的扩增引物(Artificial Sequence)
<400> 92
ttgtgttttt aactggaggc cctt 24
<210> 93
<211> 27
<212> DNA
<213> MH04KK074的扩增引物(Artificial Sequence)
<400> 93
atatttaaac aaaggctctg ggtgtaa 27
<210> 94
<211> 25
<212> DNA
<213> MH04KK074的扩增引物(Artificial Sequence)
<400> 94
cagggacttc tctagtttca tgtgt 25
<210> 95
<211> 24
<212> DNA
<213> MH05CP004的扩增引物(Artificial Sequence)
<400> 95
tgggaacaaa gtctcggatg tact 24
<210> 96
<211> 28
<212> DNA
<213> MH05CP004的扩增引物(Artificial Sequence)
<400> 96
cagcaggaca ttgacagata ctcattat 28
<210> 97
<211> 25
<212> DNA
<213> MH05CP006的扩增引物(Artificial Sequence)
<400> 97
agaaaaatgg cagagacctt gacac 25
<210> 98
<211> 28
<212> DNA
<213> MH05CP006的扩增引物(Artificial Sequence)
<400> 98
tctactttct gttctctttg tgtttccg 28
<210> 99
<211> 25
<212> DNA
<213> MH05CP010的扩增引物(Artificial Sequence)
<400> 99
caatcacatt gttccctagt gtctc 25
<210> 100
<211> 26
<212> DNA
<213> MH05CP010的扩增引物(Artificial Sequence)
<400> 100
aggtgacatt gacagagttg caaata 26
<210> 101
<211> 27
<212> DNA
<213> MH05KK020的扩增引物(Artificial Sequence)
<400> 101
aataaatcgc aatggaagca acaggaa 27
<210> 102
<211> 24
<212> DNA
<213> MH05KK020的扩增引物(Artificial Sequence)
<400> 102
ctcctagggc ttgtgagtct cata 24
<210> 103
<211> 24
<212> DNA
<213> MH05KK022的扩增引物(Artificial Sequence)
<400> 103
gttgccaatc ttaccacacc tcca 24
<210> 104
<211> 26
<212> DNA
<213> MH05KK022的扩增引物(Artificial Sequence)
<400> 104
agcctttttc ttaggacctg acatag 26
<210> 105
<211> 27
<212> DNA
<213> MH05KK062的扩增引物(Artificial Sequence)
<400> 105
tgaactgatc caacttctct ctcactg 27
<210> 106
<211> 25
<212> DNA
<213> MH05KK062的扩增引物(Artificial Sequence)
<400> 106
ctcagtgcca ttgcttatct tcctt 25
<210> 107
<211> 28
<212> DNA
<213> MH05KK078的扩增引物(Artificial Sequence)
<400> 107
caacaaaaga gaaaatctgt atagccag 28
<210> 108
<211> 28
<212> DNA
<213> MH05KK078的扩增引物(Artificial Sequence)
<400> 108
tttctgcagt tgttcatctt ctacgtta 28
<210> 109
<211> 28
<212> DNA
<213> MH05KK079的扩增引物(Artificial Sequence)
<400> 109
ttattggtct gctcagagtt tacatcag 28
<210> 110
<211> 28
<212> DNA
<213> MH05KK079的扩增引物(Artificial Sequence)
<400> 110
acagaacatt ctacccaaga ttctatgc 28
<210> 111
<211> 25
<212> DNA
<213> MH05KK122的扩增引物(Artificial Sequence)
<400> 111
caacattttt catgtggccc ctact 25
<210> 112
<211> 23
<212> DNA
<213> MH05KK122的扩增引物(Artificial Sequence)
<400> 112
ggaacaaaac aaggtgcggt ttt 23
<210> 113
<211> 24
<212> DNA
<213> MH05KK123的扩增引物(Artificial Sequence)
<400> 113
agtgttctgc cagggtcaaa ataa 24
<210> 114
<211> 25
<212> DNA
<213> MH05KK123的扩增引物(Artificial Sequence)
<400> 114
attgaatgcc aaaacctcag ggata 25
<210> 115
<211> 28
<212> DNA
<213> MH05KK124的扩增引物(Artificial Sequence)
<400> 115
cagacaagct gatctgatat ttctttag 28
<210> 116
<211> 25
<212> DNA
<213> MH05KK124的扩增引物(Artificial Sequence)
<400> 116
gccgcctaag ggatttacca atatg 25
<210> 117
<211> 26
<212> DNA
<213> MH05KK170的扩增引物(Artificial Sequence)
<400> 117
aagacctgag tagcttctgt tttctc 26
<210> 118
<211> 25
<212> DNA
<213> MH05KK170的扩增引物(Artificial Sequence)
<400> 118
ggtgctgtaa ttcccctaaa agcaa 25
<210> 119
<211> 24
<212> DNA
<213> MH06CP003的扩增引物(Artificial Sequence)
<400> 119
caaggaataa agcagtgtgt gcct 24
<210> 120
<211> 26
<212> DNA
<213> MH06CP003的扩增引物(Artificial Sequence)
<400> 120
cctcaagaat cctggaaaat gtcagc 26
<210> 121
<211> 33
<212> DNA
<213> MH06CP007的扩增引物(Artificial Sequence)
<400> 121
acactatttt aaattagtca acagttaagc ata 33
<210> 122
<211> 29
<212> DNA
<213> MH06CP007的扩增引物(Artificial Sequence)
<400> 122
ctgaaacatc actcaaaata aaaggcatt 29
<210> 123
<211> 26
<212> DNA
<213> MH06KK026的扩增引物(Artificial Sequence)
<400> 123
tctacaacta agccttttaa ccgaga 26
<210> 124
<211> 28
<212> DNA
<213> MH06KK026的扩增引物(Artificial Sequence)
<400> 124
atttcacagt tctctcttga tcatgtca 28
<210> 125
<211> 28
<212> DNA
<213> MH06KK030的扩增引物(Artificial Sequence)
<400> 125
gaatgcacag agaaattctt agaggtca 28
<210> 126
<211> 26
<212> DNA
<213> MH06KK030的扩增引物(Artificial Sequence)
<400> 126
ctccacctct tgtcttctag aaccat 26
<210> 127
<211> 28
<212> DNA
<213> MH06KK031的扩增引物(Artificial Sequence)
<400> 127
tctttgtatt cactattctt gtggctaa 28
<210> 128
<211> 25
<212> DNA
<213> MH06KK031的扩增引物(Artificial Sequence)
<400> 128
tttcaagatg ggatggagaa agcta 25
<210> 129
<211> 25
<212> DNA
<213> MH06KK080的扩增引物(Artificial Sequence)
<400> 129
ccctattcca aacctgtacc tacct 25
<210> 130
<211> 26
<212> DNA
<213> MH06KK080的扩增引物(Artificial Sequence)
<400> 130
ccccagtcac ccacctaaca tttaat 26
<210> 131
<211> 21
<212> DNA
<213> MH06KK101的扩增引物(Artificial Sequence)
<400> 131
gagcctgaga ctctgctacc a 21
<210> 132
<211> 19
<212> DNA
<213> MH06KK101的扩增引物(Artificial Sequence)
<400> 132
gggagtccca cgagcactg 19
<210> 133
<211> 28
<212> DNA
<213> MH07KK030的扩增引物(Artificial Sequence)
<400> 133
aagtgtagtc tgtgcaacaa gtttctta 28
<210> 134
<211> 27
<212> DNA
<213> MH07KK030的扩增引物(Artificial Sequence)
<400> 134
atacaaggat ttagagacca cagcatc 27
<210> 135
<211> 28
<212> DNA
<213> MH07KK031的扩增引物(Artificial Sequence)
<400> 135
ctttggagaa aactgatgag tttagctt 28
<210> 136
<211> 25
<212> DNA
<213> MH07KK031的扩增引物(Artificial Sequence)
<400> 136
cctctgtctt cttaactggc tgtag 25
<210> 137
<211> 26
<212> DNA
<213> MH07KK081的扩增引物(Artificial Sequence)
<400> 137
taagttggaa tcaccaccat tgaccc 26
<210> 138
<211> 28
<212> DNA
<213> MH07KK081的扩增引物(Artificial Sequence)
<400> 138
attcataact cctccacaca tctcagta 28
<210> 139
<211> 23
<212> DNA
<213> MH07KK082的扩增引物(Artificial Sequence)
<400> 139
tgagcttgga gcagtaaagc agg 23
<210> 140
<211> 26
<212> DNA
<213> MH07KK082的扩增引物(Artificial Sequence)
<400> 140
agtgacatca tgttgactct aactcg 26
<210> 141
<211> 28
<212> DNA
<213> MH08KK032的扩增引物(Artificial Sequence)
<400> 141
aacttgttgc agattcatgg aatcattt 28
<210> 142
<211> 26
<212> DNA
<213> MH08KK032的扩增引物(Artificial Sequence)
<400> 142
aaagagaata acagtttgac cttggc 26
<210> 143
<211> 28
<212> DNA
<213> MH09KK020的扩增引物(Artificial Sequence)
<400> 143
atgacagaag agatttctct ccagtttg 28
<210> 144
<211> 26
<212> DNA
<213> MH09KK020的扩增引物(Artificial Sequence)
<400> 144
actcgattct ttccatttcc atgtcg 26
<210> 145
<211> 25
<212> DNA
<213> MH09KK033的扩增引物(Artificial Sequence)
<400> 145
ttaaagtctc ctgtgtacac ggttg 25
<210> 146
<211> 28
<212> DNA
<213> MH09KK033的扩增引物(Artificial Sequence)
<400> 146
ctgtaccaat caagagaagt aggatgga 28
<210> 147
<211> 30
<212> DNA
<213> MH09KK034的扩增引物(Artificial Sequence)
<400> 147
gatatttgta aggtattctg gcctaaaaaa 30
<210> 148
<211> 31
<212> DNA
<213> MH09KK034的扩增引物(Artificial Sequence)
<400> 148
aagtattgaa gtgatagttt tacagtttcc t 31
<210> 149
<211> 26
<212> DNA
<213> MH09KK152的扩增引物(Artificial Sequence)
<400> 149
agacttggaa tcattcttca cagggt 26
<210> 150
<211> 25
<212> DNA
<213> MH09KK152的扩增引物(Artificial Sequence)
<400> 150
gccagaatta gcagttagca gtcat 25
<210> 151
<211> 28
<212> DNA
<213> MH09KK153的扩增引物(Artificial Sequence)
<400> 151
tttcttcctc taagtggcct cataaata 28
<210> 152
<211> 28
<212> DNA
<213> MH09KK153的扩增引物(Artificial Sequence)
<400> 152
agaattagta agctctttca cttgcagt 28
<210> 153
<211> 28
<212> DNA
<213> MH09KK157的扩增引物(Artificial Sequence)
<400> 153
actagaagca ttagaccaga ttacctgc 28
<210> 154
<211> 25
<212> DNA
<213> MH09KK157的扩增引物(Artificial Sequence)
<400> 154
acagtccatt agtgatgggt ttgtt 25
<210> 155
<211> 25
<212> DNA
<213> MH09KK161的扩增引物(Artificial Sequence)
<400> 155
cagaaaaaca gactggtcca aagtc 25
<210> 156
<211> 26
<212> DNA
<213> MH09KK161的扩增引物(Artificial Sequence)
<400> 156
cactggtttg ggaatagagt gctaag 26
<210> 157
<211> 28
<212> DNA
<213> MH10CP003的扩增引物(Artificial Sequence)
<400> 157
cccccagaaa agtatgtttt aagactct 28
<210> 158
<211> 27
<212> DNA
<213> MH10CP003的扩增引物(Artificial Sequence)
<400> 158
ccaagaccag agagataaca aatgcaa 27
<210> 159
<211> 26
<212> DNA
<213> MH10KK083的扩增引物(Artificial Sequence)
<400> 159
tttctgaatg tggcctacag tttcac 26
<210> 160
<211> 27
<212> DNA
<213> MH10KK083的扩增引物(Artificial Sequence)
<400> 160
atggaattcg aaatgatgaa gcaatga 27
<210> 161
<211> 27
<212> DNA
<213> MH10KK084的扩增引物(Artificial Sequence)
<400> 161
tgttgcttat gctgttgttc ttcaccc 27
<210> 162
<211> 27
<212> DNA
<213> MH10KK084的扩增引物(Artificial Sequence)
<400> 162
gtttgtactt ctttaaagca gggactg 27
<210> 163
<211> 25
<212> DNA
<213> MH10KK085的扩增引物(Artificial Sequence)
<400> 163
ggaggtcaag aagccttagt ttctc 25
<210> 164
<211> 25
<212> DNA
<213> MH10KK085的扩增引物(Artificial Sequence)
<400> 164
atcgtggcgc attatctctt acatc 25
<210> 165
<211> 27
<212> DNA
<213> MH10KK086的扩增引物(Artificial Sequence)
<400> 165
gcattctagc cattggacaa ttttgta 27
<210> 166
<211> 28
<212> DNA
<213> MH10KK086的扩增引物(Artificial Sequence)
<400> 166
taggtctgca ataatttccc tctactca 28
<210> 167
<211> 26
<212> DNA
<213> MH10KK087的扩增引物(Artificial Sequence)
<400> 167
actgttaagg tcaatgacgc agagta 26
<210> 168
<211> 28
<212> DNA
<213> MH10KK087的扩增引物(Artificial Sequence)
<400> 168
ttactaaagg acttggtagg tgcacata 28
<210> 169
<211> 28
<212> DNA
<213> MH10KK088的扩增引物(Artificial Sequence)
<400> 169
tttggcccat ggatagaaat aaaatgtt 28
<210> 170
<211> 28
<212> DNA
<213> MH10KK088的扩增引物(Artificial Sequence)
<400> 170
tttgaaaggc ttttgttatc aagggcta 28
<210> 171
<211> 24
<212> DNA
<213> MH10KK101的扩增引物(Artificial Sequence)
<400> 171
ccattcccta ttcagtggac tctt 24
<210> 172
<211> 24
<212> DNA
<213> MH10KK101的扩增引物(Artificial Sequence)
<400> 172
agactcagtg aggtcatgac tcaa 24
<210> 173
<211> 24
<212> DNA
<213> MH10KK163的扩增引物(Artificial Sequence)
<400> 173
gagcatcttc tccaccagtt tggc 24
<210> 174
<211> 23
<212> DNA
<213> MH10KK163的扩增引物(Artificial Sequence)
<400> 174
ttgtctcctt tcagcacaga acc 23
<210> 175
<211> 27
<212> DNA
<213> MH10KK170的扩增引物(Artificial Sequence)
<400> 175
aaagcccaca ttttgttaac atgactc 27
<210> 176
<211> 26
<212> DNA
<213> MH10KK170的扩增引物(Artificial Sequence)
<400> 176
atgtaacttc tctgaacagg gaagag 26
<210> 177
<211> 23
<212> DNA
<213> MH11CP003的扩增引物(Artificial Sequence)
<400> 177
aagcagcgat ttccatgttg ccc 23
<210> 178
<211> 23
<212> DNA
<213> MH11CP003的扩增引物(Artificial Sequence)
<400> 178
ggctgattgt ggagatgtct cct 23
<210> 179
<211> 25
<212> DNA
<213> MH11CP004的扩增引物(Artificial Sequence)
<400> 179
agaagccaaa gctccctaat agctc 25
<210> 180
<211> 27
<212> DNA
<213> MH11CP004的扩增引物(Artificial Sequence)
<400> 180
gagccagttt tgttaaagac acaatgt 27
<210> 181
<211> 27
<212> DNA
<213> MH11CP005的扩增引物(Artificial Sequence)
<400> 181
ttgctctgaa tagtgctttc agtagtg 27
<210> 182
<211> 28
<212> DNA
<213> MH11CP005的扩增引物(Artificial Sequence)
<400> 182
cagcactttc taaatagtga taggcaag 28
<210> 183
<211> 26
<212> DNA
<213> MH11KK036的扩增引物(Artificial Sequence)
<400> 183
cagctgctta tagttttgtt aagaag 26
<210> 184
<211> 25
<212> DNA
<213> MH11KK036的扩增引物(Artificial Sequence)
<400> 184
ggacccctag ataatgtcag gattg 25
<210> 185
<211> 28
<212> DNA
<213> MH11KK037的扩增引物(Artificial Sequence)
<400> 185
cttttgagat catggaaaat tccagttg 28
<210> 186
<211> 28
<212> DNA
<213> MH11KK037的扩增引物(Artificial Sequence)
<400> 186
cagaaagagg aacttaagaa gatgtggt 28
<210> 187
<211> 27
<212> DNA
<213> MH11KK038的扩增引物(Artificial Sequence)
<400> 187
ggagttctaa gcaatgagat gctaatt 27
<210> 188
<211> 26
<212> DNA
<213> MH11KK038的扩增引物(Artificial Sequence)
<400> 188
tttcccataa ttcccaaagc atggta 26
<210> 189
<211> 26
<212> DNA
<213> MH11KK039的扩增引物(Artificial Sequence)
<400> 189
agcatcattt catgcttttg aagttt 26
<210> 190
<211> 23
<212> DNA
<213> MH11KK039的扩增引物(Artificial Sequence)
<400> 190
accacctcct gtaacaacat ccg 23
<210> 191
<211> 27
<212> DNA
<213> MH11KK040的扩增引物(Artificial Sequence)
<400> 191
agaacccata gggaaacaaa ggtatgt 27
<210> 192
<211> 28
<212> DNA
<213> MH11KK040的扩增引物(Artificial Sequence)
<400> 192
tttctctcct ttcagggaac attacatc 28
<210> 193
<211> 27
<212> DNA
<213> MH11KK041的扩增引物(Artificial Sequence)
<400> 193
cattcagtat ctgtgtgcct caatgat 27
<210> 194
<211> 26
<212> DNA
<213> MH11KK041的扩增引物(Artificial Sequence)
<400> 194
ctgcagggtt ttctatccag aacaat 26
<210> 195
<211> 26
<212> DNA
<213> MH11KK089的扩增引物(Artificial Sequence)
<400> 195
cagaatgatg agctgtgcag atagcc 26
<210> 196
<211> 24
<212> DNA
<213> MH11KK089的扩增引物(Artificial Sequence)
<400> 196
gctgtctcta tgaacatccc tacc 24
<210> 197
<211> 24
<212> DNA
<213> MH11KK090的扩增引物(Artificial Sequence)
<400> 197
tgtgatggag tttatggcca acgg 24
<210> 198
<211> 25
<212> DNA
<213> MH11KK090的扩增引物(Artificial Sequence)
<400> 198
ttatgcccca aatttcactg cttag 25
<210> 199
<211> 21
<212> DNA
<213> MH11KK091的扩增引物(Artificial Sequence)
<400> 199
aactccggtc tatccaggtc c 21
<210> 200
<211> 22
<212> DNA
<213> MH11KK091的扩增引物(Artificial Sequence)
<400> 200
tgatcccatg ggactactca cg 22
<210> 201
<211> 23
<212> DNA
<213> MH11KK180的扩增引物(Artificial Sequence)
<400> 201
gcatctgagt ggctttcttc tcc 23
<210> 202
<211> 21
<212> DNA
<213> MH11KK180的扩增引物(Artificial Sequence)
<400> 202
ctgggaactt gtccggcttt a 21
<210> 203
<211> 28
<212> DNA
<213> MH11KK187的扩增引物(Artificial Sequence)
<400> 203
taggagttta tacatgatcc taagggca 28
<210> 204
<211> 26
<212> DNA
<213> MH11KK187的扩增引物(Artificial Sequence)
<400> 204
atttttggcc aaacagaatt gtttgc 26
<210> 205
<211> 21
<212> DNA
<213> MH11KK191的扩增引物(Artificial Sequence)
<400> 205
caccaaagga gctgtacctc c 21
<210> 206
<211> 24
<212> DNA
<213> MH11KK191的扩增引物(Artificial Sequence)
<400> 206
gtcaactcca aacaggcttt ttcc 24
<210> 207
<211> 26
<212> DNA
<213> MH12KK042的扩增引物(Artificial Sequence)
<400> 207
ttgcaaacta tgtcaaggac acattt 26
<210> 208
<211> 28
<212> DNA
<213> MH12KK042的扩增引物(Artificial Sequence)
<400> 208
gcaaatgatc tcagagttgc acaaattg 28
<210> 209
<211> 22
<212> DNA
<213> MH12KK043的扩增引物(Artificial Sequence)
<400> 209
gatgaacagc ttggattggg gc 22
<210> 210
<211> 25
<212> DNA
<213> MH12KK043的扩增引物(Artificial Sequence)
<400> 210
cagctgagac atagagagag gactt 25
<210> 211
<211> 26
<212> DNA
<213> MH12KK045的扩增引物(Artificial Sequence)
<400> 211
aacaggtcat ggaagcttta gatctt 26
<210> 212
<211> 27
<212> DNA
<213> MH12KK045的扩增引物(Artificial Sequence)
<400> 212
aaaatcctgg tgataaacgt acaacct 27
<210> 213
<211> 23
<212> DNA
<213> MH12KK046的扩增引物(Artificial Sequence)
<400> 213
tgtcagcttc ttgcgtgata gtg 23
<210> 214
<211> 28
<212> DNA
<213> MH12KK046的扩增引物(Artificial Sequence)
<400> 214
tttttcccca agagtctcat ctattagc 28
<210> 215
<211> 25
<212> DNA
<213> MH12KK092的扩增引物(Artificial Sequence)
<400> 215
catgtctcct tcccttggtt atacc 25
<210> 216
<211> 27
<212> DNA
<213> MH12KK092的扩增引物(Artificial Sequence)
<400> 216
aaaaattgca agagcaataa gcatgtg 27
<210> 217
<211> 25
<212> DNA
<213> MH12KK093的扩增引物(Artificial Sequence)
<400> 217
atcttttgcc ttggcatttg gtctg 25
<210> 218
<211> 27
<212> DNA
<213> MH12KK093的扩增引物(Artificial Sequence)
<400> 218
ctagtttgct tccttctatg accccta 27
<210> 219
<211> 28
<212> DNA
<213> MH12KK202的扩增引物(Artificial Sequence)
<400> 219
gagagagtga acagatgaga atcagaaa 28
<210> 220
<211> 28
<212> DNA
<213> MH12KK202的扩增引物(Artificial Sequence)
<400> 220
ttgtaatggc cttgggatca aatattct 28
<210> 221
<211> 25
<212> DNA
<213> MH13CP008的扩增引物(Artificial Sequence)
<400> 221
agagctttag taagacctca gactg 25
<210> 222
<211> 28
<212> DNA
<213> MH13CP008的扩增引物(Artificial Sequence)
<400> 222
taaaccagac tgaatgtcaa agacaaac 28
<210> 223
<211> 23
<212> DNA
<213> MH13KK047的扩增引物(Artificial Sequence)
<400> 223
gaataaccag taccaggcac ggc 23
<210> 224
<211> 25
<212> DNA
<213> MH13KK047的扩增引物(Artificial Sequence)
<400> 224
tccatccctt tgagtctatg tgtcc 25
<210> 225
<211> 28
<212> DNA
<213> MH13KK213的扩增引物(Artificial Sequence)
<400> 225
ctcttgcttc tgtcagacac ttttaatt 28
<210> 226
<211> 25
<212> DNA
<213> MH13KK213的扩增引物(Artificial Sequence)
<400> 226
cgagtctctt tttggtgtat tgcca 25
<210> 227
<211> 25
<212> DNA
<213> MH13KK217的扩增引物(Artificial Sequence)
<400> 227
ctgggaaacc agctagaaga agaga 25
<210> 228
<211> 26
<212> DNA
<213> MH13KK217的扩增引物(Artificial Sequence)
<400> 228
caaacgcact gagctattta ccttag 26
<210> 229
<211> 26
<212> DNA
<213> MH13KK218的扩增引物(Artificial Sequence)
<400> 229
gcctcccttt cagatcttac ttaggt 26
<210> 230
<211> 27
<212> DNA
<213> MH13KK218的扩增引物(Artificial Sequence)
<400> 230
aaaatgcaac acacctaata cttcagt 27
<210> 231
<211> 23
<212> DNA
<213> MH13KK225的扩增引物(Artificial Sequence)
<400> 231
atgtcaggat gctccacaac ggt 23
<210> 232
<211> 24
<212> DNA
<213> MH13KK225的扩增引物(Artificial Sequence)
<400> 232
tccacagagc atcagctatg aatc 24
<210> 233
<211> 25
<212> DNA
<213> MH13KK226的扩增引物(Artificial Sequence)
<400> 233
ctgatcttac aagttcacgg cttgt 25
<210> 234
<211> 28
<212> DNA
<213> MH13KK226的扩增引物(Artificial Sequence)
<400> 234
ttctctatat gaccagcctc tttacatg 28
<210> 235
<211> 24
<212> DNA
<213> MH14CP003的扩增引物(Artificial Sequence)
<400> 235
gctgggcata tactccaaag acag 24
<210> 236
<211> 27
<212> DNA
<213> MH14CP003的扩增引物(Artificial Sequence)
<400> 236
ccagtctcta gtaactgtcc ttctctg 27
<210> 237
<211> 30
<212> DNA
<213> MH14CP004的扩增引物(Artificial Sequence)
<400> 237
gatattagcc ctttgccaga tagataggtt 30
<210> 238
<211> 32
<212> DNA
<213> MH14CP004的扩增引物(Artificial Sequence)
<400> 238
gggaaaggat tccctattta ataaatagtg tc 32
<210> 239
<211> 22
<212> DNA
<213> MH14KK048的扩增引物(Artificial Sequence)
<400> 239
tgtctggaaa actgtagcgt gt 22
<210> 240
<211> 25
<212> DNA
<213> MH14KK048的扩增引物(Artificial Sequence)
<400> 240
ccatgcacaa ttaggaacaa cagtg 25
<210> 241
<211> 28
<212> DNA
<213> MH14KK101的扩增引物(Artificial Sequence)
<400> 241
gatgcgggat aaggaattaa tcaaggaa 28
<210> 242
<211> 27
<212> DNA
<213> MH14KK101的扩增引物(Artificial Sequence)
<400> 242
cactatgcct agctttgtct tgtctta 27
<210> 243
<211> 23
<212> DNA
<213> MH15CP001的扩增引物(Artificial Sequence)
<400> 243
gtactgcagt cacacaaagc aga 23
<210> 244
<211> 24
<212> DNA
<213> MH15CP001的扩增引物(Artificial Sequence)
<400> 244
ctaatgaaag gctgcctctg ttct 24
<210> 245
<211> 24
<212> DNA
<213> MH15CP003的扩增引物(Artificial Sequence)
<400> 245
cacacgtgct agttaggcta aata 24
<210> 246
<211> 28
<212> DNA
<213> MH15CP003的扩增引物(Artificial Sequence)
<400> 246
cttcctttgt gacttctgtt gcatttat 28
<210> 247
<211> 24
<212> DNA
<213> MH15CP004的扩增引物(Artificial Sequence)
<400> 247
cgctgtgaag tatttaacat gcag 24
<210> 248
<211> 23
<212> DNA
<213> MH15CP004的扩增引物(Artificial Sequence)
<400> 248
ggaggccttg cactgtttta tga 23
<210> 249
<211> 26
<212> DNA
<213> MH15KK066的扩增引物(Artificial Sequence)
<400> 249
tctatggatc gttcttgctt gtttct 26
<210> 250
<211> 26
<212> DNA
<213> MH15KK066的扩增引物(Artificial Sequence)
<400> 250
gggctatttt gttgactgag agaatg 26
<210> 251
<211> 28
<212> DNA
<213> MH15KK067的扩增引物(Artificial Sequence)
<400> 251
agggaaaatt cttccttatg atgggaag 28
<210> 252
<211> 30
<212> DNA
<213> MH15KK067的扩增引物(Artificial Sequence)
<400> 252
tccagtttca attttctgca cattgttaga 30
<210> 253
<211> 28
<212> DNA
<213> MH15KK069的扩增引物(Artificial Sequence)
<400> 253
tatgttgccc agaattctga gcatagac 28
<210> 254
<211> 26
<212> DNA
<213> MH15KK069的扩增引物(Artificial Sequence)
<400> 254
agggaggaaa taattcgctt tgcatt 26
<210> 255
<211> 24
<212> DNA
<213> MH15KK095的扩增引物(Artificial Sequence)
<400> 255
cagaatagca ctggatccac aggc 24
<210> 256
<211> 25
<212> DNA
<213> MH15KK095的扩增引物(Artificial Sequence)
<400> 256
aagcttaatt gccatgccgt ttatc 25
<210> 257
<211> 28
<212> DNA
<213> MH16KK053的扩增引物(Artificial Sequence)
<400> 257
gtgaagacat cgtaaaaaga tctacctg 28
<210> 258
<211> 28
<212> DNA
<213> MH16KK053的扩增引物(Artificial Sequence)
<400> 258
aatttaattg ggatcaatgc ccaaaagg 28
<210> 259
<211> 28
<212> DNA
<213> MH16KK062的扩增引物(Artificial Sequence)
<400> 259
ttattactct agaggcaggg actagcct 28
<210> 260
<211> 25
<212> DNA
<213> MH16KK062的扩增引物(Artificial Sequence)
<400> 260
aggtatctgc tgtcagtgtg actaa 25
<210> 261
<211> 27
<212> DNA
<213> MH16KK096的扩增引物(Artificial Sequence)
<400> 261
aagcatcttt ggagttctct tctccag 27
<210> 262
<211> 28
<212> DNA
<213> MH16KK096的扩增引物(Artificial Sequence)
<400> 262
tagacatatt cctacatctg tggaatgg 28
<210> 263
<211> 28
<212> DNA
<213> MH16KK255的扩增引物(Artificial Sequence)
<400> 263
ctatttcaag gtaagattct gtctctta 28
<210> 264
<211> 30
<212> DNA
<213> MH16KK255的扩增引物(Artificial Sequence)
<400> 264
aagaacatat tctaaaacag ctgaatgaac 30
<210> 265
<211> 25
<212> DNA
<213> MH16KK302的扩增引物(Artificial Sequence)
<400> 265
aatgtcattg acgtgatcac ctgca 25
<210> 266
<211> 21
<212> DNA
<213> MH16KK302的扩增引物(Artificial Sequence)
<400> 266
gtagtaggcg atgaagagcg t 21
<210> 267
<211> 25
<212> DNA
<213> MH17CP001的扩增引物(Artificial Sequence)
<400> 267
tgagttgaaa ccccagtgaa acaca 25
<210> 268
<211> 22
<212> DNA
<213> MH17CP001的扩增引物(Artificial Sequence)
<400> 268
ccccagcaat gatctcgtaa gt 22
<210> 269
<211> 24
<212> DNA
<213> MH17CP006的扩增引物(Artificial Sequence)
<400> 269
aacccttcct cctaacctca tatg 24
<210> 270
<211> 27
<212> DNA
<213> MH17CP006的扩增引物(Artificial Sequence)
<400> 270
cttacccaac agaactcaag tattggt 27
<210> 271
<211> 27
<212> DNA
<213> MH17KK014的扩增引物(Artificial Sequence)
<400> 271
tttacttaaa gcatagcttg ccttgcc 27
<210> 272
<211> 27
<212> DNA
<213> MH17KK014的扩增引物(Artificial Sequence)
<400> 272
cggttgcacc atttgacatt ctattag 27
<210> 273
<211> 24
<212> DNA
<213> MH17KK052的扩增引物(Artificial Sequence)
<400> 273
aacaggaaag cagatgaaac tggc 24
<210> 274
<211> 21
<212> DNA
<213> MH17KK052的扩增引物(Artificial Sequence)
<400> 274
ctactgtgcg tgtgcgatag c 21
<210> 275
<211> 21
<212> DNA
<213> MH17KK053的扩增引物(Artificial Sequence)
<400> 275
tggatcacaa cctcacggag g 21
<210> 276
<211> 26
<212> DNA
<213> MH17KK053的扩增引物(Artificial Sequence)
<400> 276
cgtcttggaa gtgaaaacac atcata 26
<210> 277
<211> 17
<212> DNA
<213> MH17KK054的扩增引物(Artificial Sequence)
<400> 277
gatcgcagcg gctacag 17
<210> 278
<211> 19
<212> DNA
<213> MH17KK054的扩增引物(Artificial Sequence)
<400> 278
tccatgcaca gtcccacga 19
<210> 279
<211> 28
<212> DNA
<213> MH17KK055的扩增引物(Artificial Sequence)
<400> 279
ttcataaaca agcagatatg caagaaga 28
<210> 280
<211> 25
<212> DNA
<213> MH17KK055的扩增引物(Artificial Sequence)
<400> 280
cataagccag tttcccagtt ttcaa 25
<210> 281
<211> 27
<212> DNA
<213> MH17KK077的扩增引物(Artificial Sequence)
<400> 281
ctaatgcctc tgttcaagct tctttgc 27
<210> 282
<211> 25
<212> DNA
<213> MH17KK077的扩增引物(Artificial Sequence)
<400> 282
tcaaattctt agagctccca gctga 25
<210> 283
<211> 25
<212> DNA
<213> MH17KK105的扩增引物(Artificial Sequence)
<400> 283
tttccttgga ttccacactt tgcct 25
<210> 284
<211> 25
<212> DNA
<213> MH17KK105的扩增引物(Artificial Sequence)
<400> 284
agtagatggg aaatcacacg caaat 25
<210> 285
<211> 25
<212> DNA
<213> MH17KK110的扩增引物(Artificial Sequence)
<400> 285
gcccagtaag agctttcttt tatgg 25
<210> 286
<211> 23
<212> DNA
<213> MH17KK110的扩增引物(Artificial Sequence)
<400> 286
gatgcacgct tatgggtagt gaa 23
<210> 287
<211> 21
<212> DNA
<213> MH17KK272的扩增引物(Artificial Sequence)
<400> 287
gtcttccccc aaaactgaca g 21
<210> 288
<211> 24
<212> DNA
<213> MH17KK272的扩增引物(Artificial Sequence)
<400> 288
ggactctgaa gcctctgtac acat 24
<210> 289
<211> 27
<212> DNA
<213> MH18CP003的扩增引物(Artificial Sequence)
<400> 289
cccaaaatat tactgcagat gtcctta 27
<210> 290
<211> 26
<212> DNA
<213> MH18CP003的扩增引物(Artificial Sequence)
<400> 290
agcagactaa tatgcctctg ctattt 26
<210> 291
<211> 26
<212> DNA
<213> MH18CP005的扩增引物(Artificial Sequence)
<400> 291
ctcacttttc agtattctgt tctgag 26
<210> 292
<211> 26
<212> DNA
<213> MH18CP005的扩增引物(Artificial Sequence)
<400> 292
attctgacac acaagtttat ccatgc 26
<210> 293
<211> 27
<212> DNA
<213> MH18KK285的扩增引物(Artificial Sequence)
<400> 293
ttctccttgt tcttccctgt gcatacc 27
<210> 294
<211> 27
<212> DNA
<213> MH18KK285的扩增引物(Artificial Sequence)
<400> 294
aagaagcttg aaagtctaca gttgtcc 27
<210> 295
<211> 25
<212> DNA
<213> MH18KK293的扩增引物(Artificial Sequence)
<400> 295
ctttcctccc catcaatcac ttggg 25
<210> 296
<211> 27
<212> DNA
<213> MH18KK293的扩增引物(Artificial Sequence)
<400> 296
tcaaggctat ggatacctat ctcttct 27
<210> 297
<211> 21
<212> DNA
<213> MH19CP007的扩增引物(Artificial Sequence)
<400> 297
cccagttcgg catccgtaag g 21
<210> 298
<211> 21
<212> DNA
<213> MH19CP007的扩增引物(Artificial Sequence)
<400> 298
ggtgcccaga tatggaggga a 21
<210> 299
<211> 26
<212> DNA
<213> MH19KK056的扩增引物(Artificial Sequence)
<400> 299
caactagaga tcaccccata actcag 26
<210> 300
<211> 23
<212> DNA
<213> MH19KK056的扩增引物(Artificial Sequence)
<400> 300
taaaaatgaa gattcggccg gac 23
<210> 301
<211> 26
<212> DNA
<213> MH19KK057的扩增引物(Artificial Sequence)
<400> 301
aaacagaaga gcatattggc cacaat 26
<210> 302
<211> 31
<212> DNA
<213> MH19KK057的扩增引物(Artificial Sequence)
<400> 302
gcagttaggc actaaactat attgtttcaa a 31
<210> 303
<211> 27
<212> DNA
<213> MH19KK299的扩增引物(Artificial Sequence)
<400> 303
cactccatcg tgaaagaata atcctgt 27
<210> 304
<211> 25
<212> DNA
<213> MH19KK299的扩增引物(Artificial Sequence)
<400> 304
ggttaagctg ctttgaggaa caaga 25
<210> 305
<211> 26
<212> DNA
<213> MH19KK301的扩增引物(Artificial Sequence)
<400> 305
gaatcctaag attgtggctg agagag 26
<210> 306
<211> 24
<212> DNA
<213> MH19KK301的扩增引物(Artificial Sequence)
<400> 306
gttctttcct cctgacatgg gaac 24
<210> 307
<211> 25
<212> DNA
<213> MH20KK058的扩增引物(Artificial Sequence)
<400> 307
ccaaaagtaa gaactgcttc aggga 25
<210> 308
<211> 27
<212> DNA
<213> MH20KK058的扩增引物(Artificial Sequence)
<400> 308
atgagccaca ttactttgtt ttctagg 27
<210> 309
<211> 25
<212> DNA
<213> MH20KK059的扩增引物(Artificial Sequence)
<400> 309
tgtggtgatg actgagagat gatgc 25
<210> 310
<211> 22
<212> DNA
<213> MH20KK059的扩增引物(Artificial Sequence)
<400> 310
ccatagacca gtggatgcca ac 22
<210> 311
<211> 23
<212> DNA
<213> MH20KK307的扩增引物(Artificial Sequence)
<400> 311
tgtgagtcct ctcggtcata gca 23
<210> 312
<211> 26
<212> DNA
<213> MH20KK307的扩增引物(Artificial Sequence)
<400> 312
catggcatta tcagggtctg aagaaa 26
<210> 313
<211> 24
<212> DNA
<213> MH21KK313的扩增引物(Artificial Sequence)
<400> 313
aaagcttatg tggtaggagc ctaa 24
<210> 314
<211> 27
<212> DNA
<213> MH21KK313的扩增引物(Artificial Sequence)
<400> 314
caacaagaga ggacaaattc tttcaca 27
<210> 315
<211> 26
<212> DNA
<213> MH21KK315的扩增引物(Artificial Sequence)
<400> 315
gtacctagct tagggttaga catctg 26
<210> 316
<211> 27
<212> DNA
<213> MH21KK315的扩增引物(Artificial Sequence)
<400> 316
tgtgcagaaa taacagagtg agaaagt 27
<210> 317
<211> 25
<212> DNA
<213> MH21KK316的扩增引物(Artificial Sequence)
<400> 317
gaagtccaaa gtcaaagtgt cagca 25
<210> 318
<211> 32
<212> DNA
<213> MH21KK316的扩增引物(Artificial Sequence)
<400> 318
tgttttggat gatatgtttc cttttgttca tt 32
<210> 319
<211> 24
<212> DNA
<213> MH21KK324的扩增引物(Artificial Sequence)
<400> 319
agaggagctt cacaaacatc cgct 24
<210> 320
<211> 23
<212> DNA
<213> MH21KK324的扩增引物(Artificial Sequence)
<400> 320
ctgctggtga atcagcaaaa cct 23
<210> 321
<211> 21
<212> DNA
<213> MH22KK060的扩增引物(Artificial Sequence)
<400> 321
ttatcggctg gaacgagttc a 21
<210> 322
<211> 25
<212> DNA
<213> MH22KK060的扩增引物(Artificial Sequence)
<400> 322
ggtgataaca gcttctcctg taagg 25
<210> 323
<211> 19
<212> DNA
<213> MH22KK064的扩增引物(Artificial Sequence)
<400> 323
cgtggacgcc gtgattcag 19
<210> 324
<211> 24
<212> DNA
<213> MH22KK064的扩增引物(Artificial Sequence)
<400> 324
gtgatagtgg gttttcagtg aacg 24
<210> 325
<211> 24
<212> DNA
<213> MH22KK303的扩增引物(Artificial Sequence)
<400> 325
gagccaatct tcagtcagta ccgc 24
<210> 326
<211> 22
<212> DNA
<213> MH22KK303的扩增引物(Artificial Sequence)
<400> 326
cctgtggtca cagttcttgg tc 22