msk

强生旅游官网

????–??3??”??‰?è°·???????????￠è??????o????è??è??é???????????。

??o???????-|??ˉ??é—¨??”????”??‰???o????????????????????o????????-?????o???????2??????“?????????o’?…3?3????è?¨è??è°???§????§‘?-|????¨??o???????-|??-????§‘?-|??????é?è???–°????????o????μ??o???a???????”??‰???·?????????????????è????è???2??·????-‰??‰?????o???????????ˉ????1??°?è???o??????ˉ?”¨?o???′?o?????-|èˉ??–-????a??“??–?”¨è?ˉ????ˉ????–??—…??‘?—…??o?????”??????”??‘?è°???§??o?????”????-‰é￠??????。

???è?o??ˉ??o????μ??o??????–è…èˉ′DNA?μ??o???‰è????ˉ??o???????μ??o????é????‰???é?è???μ??o???a??¨?μ????DNA????￠±??o??’??—é?o?o???????è?è§￡èˉ?DNA???é?—????ˉ??????è???????o?-￠???DNA?μ??o?????ˉ?’??”¨?o???o???/??o???????μ??o?????μ??o???a??¨?·2???????????￡?????‘?±?????|????2?‰?¤o??。

???????￡?μ??o?????ˉ?????3Sanger?μ??o?????ˉ???1977?1′???è￠?????ooèa‰??oa???o???????-|?1????a????è?±????”??‰???–?-|?????—é?·??·é???…?-??‘?????‘????o?é…??μ??o??3??????‘????μ??o??3???‰????-￡????￥?????o??μ??o?????ˉ??????è?o??o???????¨?-¤9?1′??????ABI?…??????o?o???‘????μ??o??3???¨??o?o???–?????????????°????”¨?μ??o???a???è?a?-¤?μ??o?????ˉè???…￥é￡?é???‘?±?????—???￡???????o???￡?μ??o?????ˉ??ˉ??“???????μ?????ˉ??????è|???‰Illumina?…???????Solexa?’?HiSeq????ˉ???Life Technologies???Solid????ˉ?’???—?°????454????ˉ????????‰??￡?μ??o?????ˉ??ˉè?‘????1′??”??‘??o??????????-??μ??o?????ˉ??????è|???…???Helicos?…???????????-￡???????-??μ??o?????ˉ???truesingle-moleculesequencing??? tSMSTM??‰???‰??′￥?o3?±3?-”????ˉ?…???????Oxford Nanopore Technologies?????￥????§°?‰??′￥?o3?±3?-”?…??????‰??????????-??o3?±3?-”?μ??o?????ˉ???single-molecule nanopore DNA sequencing??‰???¤a?13?′??”??‰??§‘?-|?…???????Pacific Biosciences??‰??????????-?????—??μ??o?????ˉ???single molecule real-time sequencing??? SMRT??‰?-‰??。

???2.??o????μ??o???‘?±??????2。

è????‰??￡?μ??o?????ˉ?????‰?????o??1???è§?è?¨1??‰????o”?”¨???é￠?????1?????°??????????????-¤???????￡?μ??o?????ˉ?????aè￠??·??±°???????‰?????μ??o??????o??ˉ3??￡?μ??o?????ˉ?1??-?????±é?￠??。

è?¨1.??‰??￡?μ??o?????ˉ????????o??1。

????o???￡?????o????μ??o?????ˉ????‰??·2????????o?????o????”¨????μ?????‰??￡?μ??o??°???ˉ??a??￥????¤§????‰è??????????ˉé￠?è????¨?°???￥5-10?1′??…?o?????‰??￡??o????μ??o?????…±?-????????o???￡?μ??o?????°???ˉ?μ??o??????o???????o”?”¨????μ?????‰??￡?μ??o??’??o???￡?μ??o?????ˉ”è??????????¨????????????????????ˉ??￡????1?é??????o???‰??￡?μ??o???????????o”?”¨??。

è?‘?1′??￥?…¨?????o????μ??o??????o???é??￠?é????????…¨??????è????′??￥??????????‰??μ??o??????oè§??¨??￠?é????????????ˉ?o??′2?????o????…???-??-????’???°?o|????????o?￠?é????????è?…è???o?20%?????ˉ?…¨????￠?é??????????????????é?????????—?o???°??o?”??‰????è?ˉè??????????-??????é???‘?±??????a??￥??-?????o????μ??o??????o????￠?é??????°????é￠??…¨?????。

????o?é?¨????μ??o???a??¨。

1.????¤–??o????μ??o???a。

????‰??????o???????μ??o???a??o?o?????”¨????μ??o?????ˉ????1????è|??????o??‰??￡?????°é????μ??′?o??o”?”¨è????o?1??3????è????ˉ????o???￡?μ??o???a??????è|??”??o§??????è|???‰Life Technologies???è￠?Thermo Fisher?”?è′-??‰??Illumina?’???—?°???。

1.1 Illumina?…????。

Illumina?????o?μ??o?è????????é???¤′??????????‰????????o???é￠?è??70%????????”±??????????￡??o?1998?1′?…±????????o?????é?¨????o??????????????|??°??o???￡??°?o??“￥??????è|?é”?”????é?μ??—è?ˉ?‰???2006?1′?”?è′-Solexa?…???????è?·??—?–°????￡é??é?é???μ??o?????ˉ??????è??????o????‰??????o??????????μ??μ??o?????ˉ?…??????。

Illumina?μ??o???a??ˉ?????o???é?￠???é??é?é???μ??o???a????…?????“?‰1?????o???é????o|?’?é??????????·????”??o§è§??¨??μ??o???a????…?????“?‰1?????o??o?¤§é??????’?é?????”??o§??Illumina???é?￠???é??é?é???μ??o???a??‰Miniseq?3???—a”a”MiniSeq?3???????2016?1′1?????‰???MiSeq?3???—a”a”MiSeq?3???????2011?1′2?????‰??MiSeq Dx?3???????2013?1′11?????‰?’?MiSeq FGx?3???????2015?1′1?????‰???NextSeq?3???—a”a”NextSeq500???2014?1′1?????‰?’?NextSeq550???2015?1′1?????‰??Illumina?”??o§è§??¨??μ??o???a??‰HiSeq?3???—a”a”HiSeq 2000???2010?1′1?????‰??HiSeq 2500???2012?1′1?????‰??HiSeq 3000???2015?1′1?????‰?’?HiSeq 4000???2015?1′1?????‰???HiSeqX?3???—a”a”HiSeqXFive???2015?1′1?????‰?’?HiSeqXTen???2014?1′1?????‰???NovaSeq?3???—a”a”NovaSeq 5000?3???????2017?1′1?????‰?’?NovaSeq 6000?3???????2017?1′1?????‰??。

?-¤?¤–???Illumina?μ??o???aè????…???GA?3???—a”a”GenomeAnalyzer?’?Genome AnalyzerIIx??GenomeAnalyzer???????ˉSolexa?…????è?a?·±??¨??o??????????￡?μ??o??3??????2006?1′Illumina?…?????”?è′-?o?Solexa?????¨????–-?μ??o??’??”1è‰ˉ????????¨??o?o?Genome AnalyzerIIx?’?HiSeq2000?13??°??。

Illumina?μ??o?é???”¨è?1??????è?1?μ??o?????ˉ(Sequencing by sythesis??? SBS)?’???ˉé??????ˉ?????“????ˉ???reversible terminator chemistry??‰??Illumina?…???????Genome AnalyzerIIx?’?Hiseq?o”èˉ￥èˉ′??ˉ????‰??…¨???????”¨é?????¤§???????o???￡?μ??o???o??¨???è????¤??a?3???—???????ˉ??????????????ˉ???????????è????¤??a?3???—?????o??¨é???”¨???é????ˉè?1??????è?1?μ??o?????–1?3?。

1.2??—?°??…????

?????oé????????è?ˉ???èˉ??–-??o????“????‘??￡??·¨?¤′?????—?°?é????￠?——??????èˉ??–-é?¨é—¨?????—?°?èˉ??–-??‰??ˉ??“?¤–èˉ??–-????…¨???é￠??ˉ?è…????—?°?èˉ??–-?”??‘??§‘?-|é?¨??ˉ??—?°?èˉ??–-?——?????????????o????é?¨?1??????è?a?”?è′-??·????????μ????…??????????????????￥??‰???????-??”??‰??-|??°???è??????-|?????°è?????è′¨????-|?????a??¨?’??§‘??”èˉ??‰??±??o§?“??????…???LightCycler?3???—????—?è?§?…‰???é??PCR?3????????…¨è?a??¨??·???????¤??3????MagNA Lyser?’?MagNA Pure???é???????????è??è????°??aCedex HiRes?????‘é…μ?????°?￡?μ??”??‰?è???¨?????????aCedex Bio/HT???è????‰NimbleGen?μ??o????????ˉ?é???–1?????￥???454?’???a??￥?????‰??￡???????￡?μ??o???a??。

1.2.1??—?°??μ??o??3????。

454?…??????ˉè°“?–°????￡?μ??o?????ˉ????￥???o?oo??2005?1′?o????454?…??????¨??o?o?é???‘??§?????o?o???|?￡·é…??μ??o??3????è?…é??é?é????o???????μ??o??3????a”a”Genome Sequencer 20 System?????????o?è?1??????è?1?μ??o?????…??23???1???????454?…????è￠???—?°?èˉ??–-?…??????￥1.55?o?????…??”?è′-??2007?1′?????–????????¨??o?o??§è????′??????????o???￡??o???????μ??o??3????a”a”Genome Sequencer FLX System(GS FLX)??2008?1′10???????…¨?–°???GS FLX Titanium?3???—èˉ??‰??’?è?ˉ??????è?￥?……???è??GS FLX???é?é????????-????é???o?5????????????§??èˉ?é???1?è?????-￥????????。

2015?1′?????—?°?èˉ??–-??¨??o?o?7???èˉ??–-?μ?èˉ??o§?“??’?8?????a??¨?????…???????-?èˉ??–-?o§?“?Cobas 6800?’?Cobas 8800?3?????’???????èˉ??–-?o§?“?Ventana HE 600?3????????”?è′-?o?4????…???????????????ˉAriosa Diagnostics??Signature Diagnostics??Capp Medical?’?KapaBiosystems?????oé????￠?‰“é??o????????￡?μ??o??13??°???????????????????。

2013?1′9????????—?°?????¤a?13?′??”??‰??§‘?-|?…???????PacBio??‰?-?è?￠???è???????￥PacBio???SMRT????ˉ??o??o??????‘èˉ??–-?o§?“??????…????‰??????é???¨??￠‘?’?????o”????????¨??…?????—?°???‘PacBio?”ˉ???7500???????…?????1???è??é?1???è?????PacBio?°?????‘?1????é??????′?o?????”¨????μ??o??3?????’????è—?“???。

2014?1′???????1′?????—?°??…?????”?è′-????-?èˉ??–-?…????IQuum?’??-￡??¨????‘?o3?±3?μ??o??13??°???Genia Technologies?…??????????????1′?????—?°???????Abvitro?…?????”?è′-?o??”¨?o?NGS??·?“?????¤????é????‘?ˉ?é??????ˉ?????°?o?12???????????—?°???￡????”?è′-Ariosa Diagnostics?…????????‰??……?…???“?¤–èˉ??–-?????????è??????—?????o§?‰??￡?μ????NIPT??‰è????????12???????—??????—?°??????￡????”?è′-??o?????????????…????Bina Technologies???è?????-￥??o?…???o???????-|è????′?????????è“??????。

??—?°?èˉ??–-????μ??o??????￥??‰???é???????é|–?…????è?·??—é￠?è|??§????μ??o?????ˉ????|??ˉ1?????￡?o3?±3?-”?μ??o??…????Genia????”?è′-????…?????????¨????‘?–°?13??°???????—??????′????…???¨?μ??o??????o?????°????????—?°?èˉ??–-?‰?èˉo?°??????-??????454?μ??o?èˉ??‰???è—????’???′????????????????—?????–-??¨NimbleGen SeqCap?o§?“??????¨é????o?–°?????¨??o?ˉ?é???·??????–????–°????￡?μ??o???????é????‘?ˉ?é???–1????????????????????????￥—é????????è§￡??3?–1??????è·¨è???????…?????‰?¤?é?1?”¨?o??…¨é?￠èˉ??–-???????ˉ??。

1.2.2??—?°?454?μ??o??????????。

Roche 454?μ??o??3??????ˉ???????a????????–è??è?￥?o???￡?μ??o?????ˉ????13??°???????????è|??μ??o????????|?????‰?¤o???。

???1??‰DNA?–??o“????¤?。

454?μ??o??3????????–?????????o?–1????’?illumina?????????????????ˉ????”¨?–·é???3??°???…?μ?DNA?‰“?–-???300-800bpé??????°??‰???μ????1???¨?‰???μ??¤??ˉ?????????????????￥?¤′?????–?°???…?μ?DNA????§????”¨????o¤????‰?è??è??PCR?‰??￠????è????￥è????“????????o???é“?DNA?–??o“???è§????3??‰??。

???3. DNA?–??o“????¤?。

???2??‰Emulsion PCR ????13??2PCR????…??????ˉ????a?3¨?°′??°?21???????‰1è???¨???‰。

454?μ??o???a??? DNA?‰??￠?è???¨??1??’?illumina?????a????????????????°?è???o????é“?DNA??“?????¨?°′?21??…è￠??????′????o|28um????￡???????????1???¨?…????é?￠?-μè?2??é?????。

?13??2PCR???¤§????‰1??1??ˉ??ˉ??￥??￠?????°????o??¤§?????????????o”??oé—′??￥è??è??DNA?‰??￠????…??…3é”?????ˉ??ˉa??3¨?°′??°?21a?????°′??…?21??‰?????o???è???¨???ˉ??¨PCR????o”?‰?????°???…???PCR?‰??‰????o”??????????°′?o???2?3¨?…￥??°é??é??—?è?????????‰??21è?¨é?￠????°′?o???2???é—′??￠????—???°??aè￠?????‰??21??…è￡1????°??°′??′??è???o??°??°′??′?°±???????o??????????PCR????o”??oé—′??èˉ|????-￥éa¤?|????4?‰?¤o???????3????????????ˉ???a?°??°′??′??a???????aDNA?¨?????’?????a?￡??????。

???4.?13??2PCR。

è???o?è￠??°??°′??′??…è￠?????￡????è?¨é?￠?????‰?????￥?¤′?o’è?￥???DNA?o???—???????-¤è???o????é“?DNA?o???—è???¤??‰1?????°??“?????¨?￡?????????????—??-μè?2??“?3???-?????‰PCR????o”èˉ??‰?????‰??￥???èˉ??o??ˉ???a????￡??????“???????°??‰???μé??è????????è??è??PCR?‰??￠?????1???”?‰??￠??o§?‰??????ˉ??￥??“?????°?￡???????????“????o”??????????????ˉ??￥??′????-μè?2??“?3??1??°???|??‰DNA????￡?????ˉ?é???????￥??è??è???‰??￠?????ˉ???a?°??‰???μé???°?è￠??‰??￠??o|100???????????è?è????°??????-￥?μ??o??‰è|??±????DNAé????。

???3??‰??|?￡·é…??μ??o?。

?μ??o??‰?é?è|??…??”¨???§?è?????é…??’????é“???“???è??????¤??????|??‰DNA????￡?????????￥???°??￡?????”???¨???§?PTP?13????????è???§??13???????‰1?????‰è???¤???′????o|??o44um????°??-”????ˉ???a?°??-”??…è????1?o3????a?￡???????é?è??è???§??–1?3???￥??o????ˉ???a?￡??????????????????￥????￡?μ???￥?????￥????μ??o?????o”è???¨?????|????5?‰?¤o??。

???5??|?￡·é…??μ??o?。

?μ??o??–1?3?é???”¨??|?￡·é…??μ??o??3?????°????§??ˉ”PTP???????°??-”??′?????′?°?????￡?????”??…￥?°??-”??-?????ˉ??¨?μ??o?????o”???μ??o?????o”??￥?￡????????¤§é???‰??￠???o??????é“?DNA??o?¨????????ˉ????????o”????…￥???§?dNTPè??è????????????o”???|????dNTPè???????…?μ??o???—é…??ˉ1???????????¨?????????é???”???|?￡·é…???o??￠??é???”??????|?￡·é…???o??￠??????????o”??“?3???-???ATP???é…???–?-|é…?????o”?”????ATP???”???????ATP?’?è?§?…‰?′?é…??…±????°§??–????μ??o?????o”??-???è?§?…‰?′?????-??1???‘??oè?§?…‰???????—??”±PTP?????|????§???CCD?…§?????oè?°???????????é?è??è????—??oè??è???…‰?????·?¤????è?è?·??—?????????μ??o???“??????”±?o??ˉ????§?dNTP??¨????o”??-?o§?”????è?§?…‰é￠?è‰2?????????????-¤??ˉ??￥??1???è?§?…‰???é￠?è‰2??￥??¤?–-è￠??μ?????-?????o???—??????o”??“?????????????|????dNTP?????¨????￡·é…?é…????????”¨???é??è§￡ATP??????è??ˉ?è?′è?§?…‰?·???-?????￥???????μ??o?????o”è???…￥???????a??a??ˉ???”±?o?454?μ??o?????ˉ??-????ˉ???a?μ??o?????o”é????¨PTP????????????????°??-”??-è??è????????è?è???¤§?¤§é?????????o’é—′????12?‰°?’??μ??o?????·???。

454????ˉ???¤§???????????¨?o??…?è??è?·??—è??é??????μ??o?èˉ?é???????“?‰?454????ˉ????13???èˉ?é????ˉè??400bp????1???”454????ˉ?’?illumina???Solexa?’?Hiseq????ˉ?????????????????è|????????a??o??1??ˉ?—??3????????μ?é?????è???‰????é???o|????|???“?o???—??-?-???¨?±?????o?PolyA?????…??μ?—?????μ??o?????o”????????????…￥?¤???aT???è??‰????…￥???T?????a??°??aè??é?è??è?§?…‰??o?o|??¨?μ?è?·??—???è???°±??‰??ˉè???ˉ?è?′??“???????????????1??-￡??ˉ?”±?o?è?????????????454????ˉ?????¨?μ??o?è???¨???-????…￥??’?…￥?’???o?¤±????μ??o?é”?èˉˉ??。

1.3 ABI?…????/Life Technologies。

2008?1′???Life Technologies?”±??????Invitrogen?…?????’??o”?”¨?”??‰??3?????…???????applied Biosystems??????§°ABI??‰????1?è???????é???????¨2013?1′4??????èˉ￥?…???????è￠?èμ?é??é￡???–?°”?§‘???…???????Thermo Fisher Scientific??????§°èμ?é??é￡???–?°”??‰?”?è′-??。

èˉ′èμ·?…¨è?a??¨?μ??o???a???ABI?…????é?￡??ˉ????ˉ1???é￠??¤′???????…?3730?——è?°?μ??o???a??¨?oo?±???o??????è????’??-??????èμ?èμ????????????è??????¨?–°????￡?μ??o??–1é?￠???454?’?Solexa??￠?o??…???o???????…???¨??o?o?NGS??a??¨??ABI??“?????????????￡????è§????è????￠??·??????éa???¤é???‘?????ˉ1?‰??????a??¨???o???ˉ????????¨2006?1′?–￥èμ?1.2?o?????…??”?è′-?o?é?—??????????…????Agencourt Personal Genomics???APG??‰?????o?????ˉ????o??1′?°??–°????￡?μ??o??3??????¨??‘?????o??APG?????????????ˉ??ˉSOLiD???supported oligo ligation detetion??‰????…?????‰1?1??¤???¨?o???￥???è‰2è?§?…‰???è?°?ˉ????è?·é…????è????-è????￥????????o??o???????–??￡?o??????????è?????é…?è????￥????o”?????ˉ?ˉ1?????·è′?DNA?‰???μè??è???¤§è§??¨??‰??￠??’?é??é?é???1?è???μ??o???。

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