utb 8449: thomas reinard, molekularbiologische methoden - leseprobe
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7/23/2019 UTB 8449: Thomas Reinard, Molekularbiologische Methoden - Leseprobe
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]TF 2448
Mhom Kvfmhtycmamhoyibket dmv Umvjkcm
Fbjk} Umvjkc Ljo Smhakv ShmoUmvjkc Fkvfkvk F}dvhib G~jkdmo Ekvahoctgo Bhjjyeki}jtky/s}u ShmoShjbmja Ehol AoibmoK/ Evkoilm Umvjkc Tfhocmo }od FkymjBk}~t Umvjkc Fmvo Yt}ttckvt Shmo@}jh}y Ljholbkvdt Umvjkcyf}ibbkodj}oc Fkd Bmhjfv}ooJ}ih}y % J}ih}y Umvjkcycmymjjyibket Yt}ttckvtAgbv Yhmfmil TfhocmoOgagy Umvjkcycmymjjyibket Fkdmo!FkdmoGvmjj Eyyjh Umvjkc PvhibMvoyt Vmhobkvdt Umvjkc Aoibmo FkymjEmvdhokod Yibohocb ^kdmvfgvo Aoibmo Shmo PvhibM}cmo ]jamv Umvjkc Yt}ttckvt]UL Umvjkcycmymjjyibket LgoytkopUkodmobgmil % V}~vmibt Ctthocmo Gkluhjjmude Bgibyib}jumvjkc KC ko dmv MTB Pvhib
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Tbgaky Vmhokvd
AjmljkfhjchibmAmbdmo
;63 Kffhjd}ocmo;= Ak~y4; Tkfmjjmo
Umvjkc M}cmo ]jamv Yt}ttckvt
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4 Ha~vmyy}a
Fhfjhgcvkehyibm Hoegvakthgo dmv Dm}tyibmo Okthgokjfhfjhgtbml
Dhm Dm}tyibm Okthgokjfhfjhgtbml umvpmhibomt dhmym }fjhlkthgomo ho dmv Dm}tyibmo Okthgokjfhfjhgcvkm;> M}cmo ]jamv LC
Sgjjcvkysmc 4;* 3>=88 Yt}ttckvt "Bgbmobmha(
M!Akhj0 hogN}jamv/dm
Hotmvomt0 sss/}jamv/dm
Jmltgvkt0 Kjmyykodvk Lvmhfk}a
Bmvytmjj}oc0 @vcmo Y~vmopmj
]ayibjkcmots}v* Hoomojkxg}t }od Yktp0 Ktmjhmv Vmhibmvt* Yt}ttckvt
Dv}il }od Fhod}oc0 Mcmdyk* Ykfkdmjj* Y~kohmo
^vhotmd ho Y~kho
HYFO 832!9!26=6!2448!3 "]TF!Fmytmjjo}aamv(
Tbgaky Vmhokvd*s}vdm ;859 ho Jmumvl}ymo cmfgvmo }od yt}dhmvtm okib dma Kfht}v ;826 ko dmv
Evhmdvhib!Shjbmjay!]ohumvyhtt Fgoo Fhgjgchm/ Dky Tbmak ymhomv Dhyymvtkthgo skv dhm Ibkvkltmvhyhmv}oc
ugo K}rho!fhodmodmo ^vgtmhomo }od s}vdm ka Hoytht}t v Cmomthl }od mhoma jocmvma K}motbkjt ko
dmv ]ohumvyhtx g Ogvtb Ikvgjhok kt Ibk~mj Bhjj d}vibcmbvt/ Koyibjhmmod smibymjtm mv ko dky Hoytht}t
v ^jkopmocmomthl* Kft/ HH* dmv Jmhfohp ]ohumvyhtt Bkoogumv "btt~0''sss/cmomthl/}oh!bkoogumv/dm(* sgmv kjy Klkdmahyibmv Vkt kvfmhtmt/ Dhm ugo hba cmjmhtmtm Kvfmhtycv}~~m Agjml}jkvm Fhgibmahm fmyibthct
yhib aht dmv Bmvytmjj}oc ~bkvakpm}thyib vmjmukotmv ^vgtmhom ho ^ekopmo ygshm aht dmv Shvl}oc ugo K}rhomo/
Mv fhmtmt ho dmo Yt}dhmococmo Jhm Yihmoim* ^jkopmofhgtmibogjgchm }od Fhgjgchm mhcmoumvkotsgvtjhib
Jmbvumvkoytkjt}ocmo ho dmo Eibmvo Agjml}jkvfhgjgchm }od Fhghogvakthl ko/
Mv hyt mhomv dmv K}tgvmo dmv Dmibmak!Yt}dhm Fhgtmibogjgchm 6>6> "btt~0''fhg6>6>/dmibmak/dm(/
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=
; Dk Jmfmo od mhom Fmkod!mhjm ;;
;/; Dmv K}fk} ugo DOK }od VOK ;9;/6 Dmv cmomthyibm Igdm ;5;/9 Dhm Cmom ;3;/4 Dhm ^vgtmhom 66;/= Dhm Pmjjm 63
6 Codjkcmo dm Kfmh ha Jkf 68
6/; Skyymv 9>6/6 Amyy}oc dmy ~B!Smvtmy 9;6/9 ^}mv 966/4 Skkcmo 996/= Ahlvg~h~mttmo 946/5 Cmm ha Jkfgv 95
6/3 Pmotvh}cmo 956/2 Ahyibmo }od Lgopmotvhmvmo 4>6/8 Lgopmotvhmvmo 4;6/;> ^}mvsmibymj 466/;; ^vgfmo!Jkcmv}oc 466/;6 Ytmvhjmy Kvfmhtmo 496/;9 Dhm ljmhomo Evm}odm ^emcm }od
K}p}ibt ugoMyibmvhibhk igjh 4=6/;4 Dmv K}yibj}yy ugo Cmsmfmo =>
9 Kemhohcoc uo Oljmhomo =9
9/; Cmcmoy~hmjmv mvgjcvmhibmv DOK! }odVOK!Hygjkthgomo ==
9/6 Mrtvklthgo ugo O}ljmhoy}vmo =29/9 Dhm smhtmvm K}vmhohc}oc dmv DOK =89/4 Mhohcm k}ycmsbjtm
DOK!K}vmhohc}ocyamtbgdmo 559/= Dhm Hygjkthgo ugo VOK 3;9/5 fmvyhibt fmv dmo Mhoyktp ugo Lhty 349/3 Th~~y p}a Mvphmjmo bgbmv K}yfm}tmo
fmh dmv Hygjkthgo ugo O}ljmhoy}vmo 349/2 [}kothphmv}oc ugo O}ljmhoy}vmo 3=
4 ^jxamkm Lmmomklho 38
4/; ^vhoph~ dmv ^gjxamvkym!Lmttmo!vmklthgo 2;
4/6 Dhm Lga~gomotmo dmv ^IV 2;4/9 Cmvtm v dhm ^IV 8>4/4 Dhm Ytkodkvd!^IV 8;4/= K}ycmsbjtm ^IV!Amtbgdmo 8;4/5 Kosmod}ocmo dmv ^IV ;>>4/3 G~thahmv}oc dmv ^IV!Vmklthgo ;>;
= Ljohmmo e Mhomhcm ;>9
=/; Vmytvhlthgoymopxam ;>==/6 Jhckthgo ;;9=/9 Dm~bgy~bgvxjhmv}oc ;;==/4 Dhm Tvkoygvakthgo ugoM/ igjh ;;3
=/= Dmv Smc p}a ljgohmvtmo Cmo ;;8=/5 Dmv }ocmvhibtmtm Mhofk} mhomv
ka~jhphmvtmo DOK ho mhomo Umltgv ;;8=/3 Dmv cmvhibtmtm Mhofk} ugo DOK
ho mhomo Umltgv ;66=/2 Jhckym!}okfbochcm
Ljgohmv}ocyyxytmam ;64=/8 Smoo my akj ohibt ljk~~t /// ;6=
5 Umlmo ;63
5/; ^jkyahdm ;685/6 Ljgohmv}ocy~jkyahdm ;965/9 Vmlgafhokym!fkyhmvtm Ljgohmv}oc ;955/4 Mr~vmyyhgoy~jkyahdm ;985/= Vm~gvtmvcmom ;495/5 ^bkcmo ;455/3 ^bkcmahdm ;435/2 Yb}ttjm!Umltgvmo ;435/8 Loytjhibm Ibvgagygamo0 FKIy*
^KIy }od XKIy ;425/;> Bmmo kjy Ljgohmv}ocy! }od
Mr~vmyyhgoyyxytma ;48
Hobkjummhiboh
Ugvsgvt 3
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5 Hobkjtyumvpmhibohy
5/;; Dhm Tvkoygvakthgo ugo ^ekopmo ;=;5/;6 Dhm Tvkoygvakthgo ugo Y}cmthmvmo
}od thmvhyibmo Pmjjl}jt}vmo ;=9
3 Mjml~bmm uoOljmhomo ;=3
3/; Kckvgym!Cmjmjmltvg~bgvmymugo DOK ;=8
3/6 Dhm Dmtmlthgo dmv DOK ha Cmj ;543/9 ^v~kvkthum Kckvgymcmjm ;5=3/4 K}tvmoomo ugo VOK ha
Kckvgymcmj ;533/= Embjmvy}ibm fmh dmv Kckvgym!
Cmjmjmltvg~bgvmym ;52
2 ^mhokemhohcoc ;3;
2/; Bgagcmohykthgo ;3=2/6 Mrtvklthgo ugo ^vgtmhomo ;332/9 Dhkjxym }od Lgopmotvhmv}oc ;2;2/4 Smhtmvm K}vmhohc}ocyyibvhttm ;292/= [}kothphmv}oc ugo ^vgtmhomo ;252/5 K}vmhohc}ocyumvkbvmo v
cmtkcctm ^vgtmhom k}yM/ igjh ;28
8 Cmjmjml~bmm uo^mhomo ;8=
8/; Dhm ^gjxkivxjkahd!Cmjmjmltvg~bgvmym"^KCM( ;83
8/6 Dhm dmokt}vhmvmodm YDY!^KCM ;838/9 Dky Evfmo dmv Cmjm 6>58/4 Smhtmvm Mjmltvg~bgvmymumvkbvmo
v ^vgtmhom 6>2
8/= Smytmvo Fjgtthoc 6;;8/5 Smoo my akj ohibt ljk~~t /// 6;4
;> HaaofhibmahibmAmbdmo 6;3
;>/; Kothlv~mv 6;8
;>/6 ^vhoph~hmo haa}ofhgibmahyibmvUmvkbvmo 669
;>/9 Y~mphyibm }od }oy~mphyibmFhod}ocmo 668
;>/4 Haa}ofhgibmahyibm Umvkbvmo 69>;>/= MJHYK 69>;>/5 Haa}ovf}oc okib Smytmvo Fjgt 69=;>/3 Haa}ogkohttyibvgaktgcvk~bhm 693;>/2 Smhtmvm Kothlv~mv!fkyhmvtm
Amtbgdmo 698;>/8 Smoo my akj ohibt ljk~~t /// 64>
;; Ajmljkfhjchm eEcmibhmom 649
;;/; DOK!Ym{}mophmv}oc 64=;;/6 Fhghogvakthyibm Ym{}mopkokjxym 642;;/9 Dmv Mhoyktp dmv Fhghogvakthl v
dhm Ljgohmv}oc ugo DOK 6=>;;/4 Ugjjytodhcm Ym{}mopmo d}vib
9 VKIM!^IV }od = VKIM!^IV 6=;
;;/= Ytkaaykaaj}ocmo 6=9;;/5 Hdmothphmv}oc ugo O}ljmhoy}vmo
d}vib Bxfvhdhyhmv}oc 6=9;;/3 DOK!Ibh~y gdmv Ahivgkvvkxy 6==;;/2 Umvodmv}oc ugo O}ljmhoy}vmo 6=5;;/8 Yxotbmthyibm Cmom 65>
Kobkoc
K; Yhibmvbmht ha Jkfgv 656
K6 Dhm ;> cgjdmomo Jkfgvvmcmjo 659K9 Dky Jkfgvf}ib }od dhm okjm Kvfmht 654
Umvpmhibohy dmv Ak~y* ^vgtglgjjm* M!Dgiy*Tkfmjjmo }od Kffhjd}ocmo 655Kflvp}ocyumvpmhibohy 652[}mjjmoumvpmhibohy 652Vmchytmv 658
Kjjm M!Dgiy odmo Yhm ha Hotmvomt }otmv btt~0''sss/}tf!ambv!shyymo/dm
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Dhmymy F}ib fkyhmvt k} amhomv cjmhibokahcmoUgvjmy}oc* dhm yhib ko gvtcmyibvhttmom Yt}dhm!vmodm dmv Jmfmoyshyymoyibktmo vhibtmt/ My ygjjdhm Jilm pshyibmo dmo kjjcmamhomo Jmbvf!ibmvo dmv mvytmo Ymamytmv }od vmhomo Amtbgdmo!fibmvo yibjhmmo/Dkvfmv bhok}y hyt my mho Vmhymbvmv d}vib dkyp}oibyt }omotdmiltm Jkod agjml}jkvfhgjgchyibcm~vctmv Jmfmoyshyymoyibktmo aht kjj hbvmo Ek!imttmo/ Dkbmv shvd Hbomo dhmymy F}ib k}ib y~tmvha Jkfgv mho bgmotjhib smvtugjjmv Fmvktmv ymho/Dmoo gfsgbj dhm Kvfmht ha Jkfgv bm}tm d}vibdhm Umvsmod}oc ugo Lhty fmythaat hyt* motbmfmodhmym dmo mr~mvhamothmvvm}dhcmo Yt}dhmvmodmoohibt ugo dmv Ogtsmodhclmht* dhm p}cv}odmjhm!cmodmo ^vhoph~hmo p} lmoomo/ O}v yg lkoo dmvEmbjmv cm}odmo smvdmo* smoo mho Umvkbvmo
ohibt shm ugvcmymbmo }olthgohmvt/ Ogib uhmjshibthcmv hyt my `mdgib* dkyy dky Embjmo dhmymvLmootohyym dkp} bvmo lkoo* Mvcmfohyym kjyibp} hotmv~vmthmvmo gdmv dkyy shibthcm Lgotvgjjmoumvcmyymo smvdmo/Dhmymy F}ib lkoo o}v fmhy~hmjbkt dhm cvgm Pkbjdmv agjml}jkvfhgjgchyibmo Amtbgdmo fmjm}ib!tmo* dkbmv s}vdm dmv Egl}y k} dhm Klt}kjhtt}od Umvfvmht}oc dmv Umvkbvmo cmymtpt/ Gt jkyymoyhib fmh }otmvyibhmdjhibmo Amtbgdmo ytkvlm ^k!
vkjjmjmo k}yakibmo* smybkjf dhm Igoim~t!Ak~ydmv p}oibyt umvshvvmodmo Uhmjkjt dmv Umvkb!vmo mhom Ytv}lt}v cmfmo }od P}ykaamobocmpshyibmo hbomo k}dmilmo/ Dmv Tmrt shvd d}vibumvyibhmdmom Tmrtfgrmo k}cmjgilmvt0
C } t p } s H y y M O !Fgrmo jhmmvo Bho!tmvcvodm/
t H ^ ^ !Fgrmo cmfmo smvtugjjm Th~~y vdhm ~vklthyibm Kvfmht/
^ v g t g L g J J !Fgrmo umvugjjytodhcmofmygodmvy shibthcm Umvkbvmo aht p}ytp!jhibmo }od dmtkhjjhmvtmo Hogvakthgomo/
]a dky F}ib ~vmhyjhib mvyibshocjhib p} bkjtmo*a}yytmo jmhdmv mhohcm hotmvmyykotm Amtbgdmo k}y!cmjkcmvt smvdmo }od ytmbmo dkbmv o}v gojhom p}vUmvc}oc/ My jgbot yhib ymbv* ho dmo }akocvmh!ibmo Gojhom!Fmvmhib ambv kjy o}v mhomo Fjhil p}smvmo/Oktvjhib motytmbt mho F}ib ohibt ha ythjjmo La!amvjmho/ Gbom dhm Bhjm ugo amhomo Lgjjmcmo}od Evm}odmo svm dky ugvjhmcmodm Smvl yhibmv}omvthc dkbmv cmlgaamo* dkbmv aibtm hibKodv Evmopmj* Shmflm Vktb`m }od Akvmo Shib!akoo ckop bmvpjhib v dhm uhmjm lgoytv}lthumoDhyl}yyhgomo }od Ugvyibjcm dkolmo/ AkvthokEka}jjk bkt dhm Mvcmfohyym* dhm ho uhmjmo Egtgyp} ymbmo yhod* ha Jkfgv ~vgd}phmvt/ Amhomo Aht!kvfmhtmvo }od Lgjjmcmo* dhm hbvm Egtgy p}v Umv!c}oc cmytmjjt bkfmo* aibtm hib mfmokjjy bmvp!
jhib dkolmo/Gbom amhom Ekahjhm* Mjuhvk* Bmovhl }od Ijkvk*dhm ahib dhm ckopm Pmht }otmvyttpt bkt* svmdhmymy ^vg`mlt ohmakjy umvshvljhibt sgvdmo/Dkolm+Yg* }od dhmy ygjj cmo}c dmv Ugvvmdm ymho/ Ytmhcmoshv o}o mho ho dhm Smjt dmv agjml}jkvfhgjgch!yibmo Amtbgdmo* kfmv ohibt gbom mhomo jmtptmoshibthcmo Bhosmhy p} cmfmo0Yhibmv shvd dhm mho gdmv kodmvm bhmv fmyibvhm!
fmom Amtbgdm ho akoibmo Jkfgvmo kfsmhibmodd}vibcmbvt/ Smoo dky ^vgtglgjj }olthgohmvt*chft my lmhomvjmh Cv}od dkvko mtsky p} odmvo/Fmugv shv }oy o}o k} dmo y~koomodmo Smcd}vib dhm Amtbgdmo dmv Agjml}jkvfhgjgchm fm!cmfmo* fjmhfmo ahv o}v ogib psmh Sgvtm ahtp}!tmhjmo0
LMHOM ^KOHL+
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K} dmo mvytmo Fjhil yibmhot dhm Agjml}jkvfhg!jgchm k}y mhoma }ofmvyibk}fkvmo Ykaamjy}!vh}a ugo Amtbgdmo p} fmytmbmo* sky dhmymoFmvmhib dmv Okt}vshyymoyibkt pskv y~koomodkfmv k}ib ymbv }ofmvyhibtjhib akibt/ Mhcmot!jhib hyt my kfmv ckv ohibt yg yibsmv* dmoo cjil!jhibmvsmhym jhmcmo umvyibhmdmomo Amtbgdmo gtbojhibm ^vhoph~hmo p}cv}odm/Ak~ ; pmhct mhomo fmvfjhil dmv ho dhmyma F}ibfmbkodmjtmo Umvkbvmo/ Shv fmchoomo ha Pmot!
v}a* fmh dmo Fk}ytmhomo dmy Jmfmoy "Lk~htmj ;(/Okib mhomv l}vpmo Fmyibvmhf}oc cochcmv Jkfgv!cmvtm "Lk~htmj 6( fmyibthcmo shv }oy aht dmvK}vmhohc}oc dmv Fhgagjmljm* sgfmh shv }oyp}oibyt k} dhm DOK! }od VOK!K}vmhohc}oclgopmotvhmvmo "Lk~htmj 9(/Dhm gjxamvkymlmttmovmklthgo "^IV* Lk~htmj 4( hytmho pmotvkjmv Dvmb! }od Kocmj~}olt/ Yhm ytmjjt dmofmvckoc p} dmo umvyibhmdmomo Amtbgdmo dmvLjgohmv}oc }od Tvkoygvakthgo dkv "Lk~htmj =(/
p dhmma Fib
K}evmhohc}oc
Jmfmsmymo
MJHYK
VOK CmjOgvtbmvo Fjgt
Bxfvhydhyhmv}oc
Vmumvym
Tvkoylvh~thgo
^IV
Kfjmhtmo ugo vhamvok}y dmv Ym{}mophmv}oc
Mjmltvg~bgvmymSmytmvo Fjgt
Haa}oevf}oc
BgagcmohykthgoMrtvklthgoEjj}ocmo
BgagcmohykthgoMrtvklthgoEjj}ocmo
BgagcmohykthgoMrtvklthgoEjj}ocmo
Ibvgaktgcvk~bhm ^vgtmhom VOK
DOK
DOK Ym{}mop
^vgtmhoym{}mop
Fhghoegvakthl Ljgohmv}oc
}odTvkoyegvakthgo
Ak~ ;0fmvyhibt dmv mhopmjomo Kvfmhtyyibvhttm ho dmv Agjml}jkvfhgjgchm* smjibm ho dhmyma F}ib fmbkodmjt smvdmo/
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8P} dhmyma F}ib
Ho dhmyma P}ykaamobkoc smvdmo shv }oy k}ibaht dmo Umltgvmo fmyibthcmo "Lk~htmj 5(/ Dkyyhod ymjfyt!vm~jhphmvmodm DOK!Agjmljm* ho dhmmho vmadmy DOK!Evkcamot mhocmymtpt smvdmolkoo* dkaht yhm my ho mhom om}m Shvtypmjjm fvho!cmo/ Dhm fmlkootmytmo Umltgvkvtmo yhod ^jky!ahdm/ K} dhmym Smhym motytmbmo tvkoycmom Gv!ckohyamo* smjibm d}vib dhm K}okbam vmadmvDOK om}m Mhcmoyibktmo mvbkjtmo bkfmo* fmh!y~hmjysmhym ytmjjmo yhm mho om}my ^vgtmho bmv/Mhohcm dhmymv Gvckohyamo yhod v agjml}jkv!fhgjgchyibm Kvfmhtmo fmygodmvy c}t cmmhcomt}od smvdmo kjy Agdmjjgvckohyamo fmpmhibomt/^vgd}phmvt mho tvkoycmomv Gvckohya}y mho vma!dmy ^vgtmho* y~vhibt ako ugo bmtmvgjgcmv Mr!~vmyyhgo/ Dhm K}vmhohc}oc }od Kokjxym ygjibmvbmtmvgjgcmv ^vgtmhom hyt Cmcmoytkod dmv Lk~h!
tmj 2 }od 8/ Dmv Okibsmhy fmythaatmv ^vgtm!hom fmyibvolt yhib bhmv k} Kothlv~mv!fkyhmvtmOkibsmhyumvkbvmo shm Haa}ovf}oc }odMJHYK "Lk~htmj ;>(/ Ho Pmhtmo dmv Cmoga~vg`mltmohaat dhm Hogvakthgoyy}ibm fmv dky Hotmvomtmhomo smymotjhibmo Tmhj dmv Kvfmhtypmht mhomyAgjml}jkvfhgjgcmo ho Koy~v}ib* smybkjf ho Lk!~htmj ;; p}ahodmyt mho l}vpmv Mhofjhil ho dhmymoTbmamofmvmhib p} odmo hyt/ Ho dhmyma Lk~htmj
fmyibthcmo shv }oy k}ib aht mhohcmo umvfvmh!tmtmo Umvkbvmo* dhm cmshyym Ugvlmootohyym mv!gvdmvo* smjibm p}ugv ho dmo kodmvmo Lk~htmjoumvahttmjt s}vdmo/P} dhmyma F}ib cmbvt k}ib mho }akocvmhibmvGojhom!Fmvmhib* ho dma Yhm ygsgbj ^vgtglgjjm kjyk}ib Hogvakthgomo p} Tmibohlmo odmo* dmvmoAmtbgdhl dmo Vkbamo dhmymy F}ibmy y~vmocmosvdm/ Bm}tm chft my uhmjm ymbv otpjhibm Umvkb!vmo* dhm smcmo hbvmy k~~kvkthumo K}skody gdmvdmv Lgytmo mbmv ho Akytmv! gdmv Dgltgvkvfmhtmomhocmymtpt smvdmo/ Ygjibm klt}mjjm Tmibohlmoodmo yhib mfmokjjy ha Gojhom!Fmvmhib p} dhm!yma F}ib/Mhomo Bhosmhy aibtm hib smcmo ymhomv Shibthc!lmht ko dky Modm dhmymv Mhojmht}oc ymtpmo0Uhmjm Amtbgdmo }otmvyibmhdmo yhib ho hbvmv
D}vibbv}oc ugo Jkfgv p} Jkfgv/ Dhmy fmdm}tmt*dkyy mhom ho dhmyma F}ib fmyibvhmfmom Amtbgdmambv gdmv smohcmv ytkvl umvodmvt d}vibcmbvtsmvdmo lkoo/ Dkbmv yhod dhm k}cmjhytmtmo ^vg!tglgjjm kjy Ugvyibjkc p} fmtvkibtmo }od ohibt kjyytvhltm Kvfmhtykojmht}oc/ Yhm s}vdmo k}cmoga!amo }a dhm p}cv}odmjhmcmodmo ^vhoph~hmo p}umvdm}tjhibmo* dmoo jmtptmodjhib pbjt o}v mhomYkibm0 Mhom Amtbgdm ygjjtm }olthgohmvmo/
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;;
; Dk Jmfmood mhom Fmkodmhjm
;/; Dm Kefk uo DOK od vOK
;/6 Dm cmomhibm Idm
;/9 Dhm Cmom
;/4 Dhm ^mhom
;/= Dhm pmjjm
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Dky Jmfmo }od ymhom Fmytkodtmhjm;6
K} dmo mvytmo Fjhil mvyibmhot dky Jmfmo k} dmvMvdm mrtvma uhmjjthc/ Yibgo ho dmv ljmhoytmofhgjgchyibmo Mhobmht* dmv Pmjjm* odmo shv lga!~jmrm ytv}lt}vmjjm }od }olthgokjm P}ykaamo!
bocm/ K} dmo psmhtmo Fjhil jkyymo yhib kjjcm!amhom ^vhoph~hmo motdmilmo* dhm my mvjmhibtmvo*Cmamhoykalmhtmo pshyibmo dmo uhmjjthcmo fhg!jgchyibmo Ugvcocmo p} motdmilmo0;/ Kjjm fhgjgchyibmo Yxytmam yhod bhmvkvibhyib
ytv}lt}vhmvt/6/ My chft ho fhgjgchyibmo Yxytmamo mhom ymbv
mocm Umvlo~}oc pshyibmo dmv Ytv}lt}v }oddmv E}olthgo/
Cmvkdm mhoakj uhmv umvyibhmdmom Kvtmo ko Ak!lvgagjmljmo yhod v dhm mogvam Uhmjkjt umv!kotsgvtjhib/ Ho dhmyma F}ib fmkyymo shv }oyaht dmo O}ljmhoy}vmo }od dmo ^vgtmhomo/ ^g!jxykiibkvhdm }od Jh~hdm ytmjjmo mfmokjjy shib!thcm Aklvgagjmljm dkv* smvdmo bhmv `mdgibohibt fmbkodmjt/Kjjm Aklvgagjmljm yhod bhmvkvibhyib k}cm!fk}t* dmoo yhm fmytmbmo k}y Agogamvmo* kjygljmhomvmo Agjmljmo/ Dhm Agogamvm smvdmofmv mhom Lgodmoykthgo }otmv Kfy~kjt}oc ugoSkyymv p} haamv cvmvmo Mhobmhtmo p}yka!
amocmct0 O}ljmgthdm smvdmo p} O}ljmhoy}!vmo lgodmoyhmvt* Kahogy}vmo p} ^m~thdmo }od^vgtmhomo/ Ygjibm Aklvgagjmljm loomo d}vibBxdvgjxym shmdmv ho ljmhomvm Mhobmhtmo* fhy bhop} dmo Agogamvmo pmvjmct smvdmo/Bm}tm hyt kjjcmamho klpm~thmvt* dkyy Vhfgo}ljm!hoy}vmo "VOK( dhm jtmytmo Fhgagjmljm dkv!ytmjjmo/ Mhomvymhty loomo yhm ho hbvmv Ym{}mopcmomthyibm Hogvakthgomo y~mhibmvo* shm dhmy
ho VOK!Uhvmo dmv Ekjj hyt* kodmvmvymhty yhod yhmk} uhmjjthcm Smhym mopxakthyib klthu* shm hoVhfgygamo gdmv Y~jhimgygamo/ Y~tmv ho dmvMugj}thgo bkfmo yhib kodmvm Fhgagjmljm k}
`msmhjy mhom dmv fmhdmo E}olthgomo y~mphkjhyhmvt/Dhm Dmygrxvhfgo}ljmhoy}vm "DOK( hyt ibmahyibuhmj ytkfhjmv }od }ochmvt kjy Y~mhibmv dmv cmomth!yibmo Hogvakthgo* ^vgtmhom dhmomo ygsgbj dmvYtv}lt}vfhjd}oc* kjy Y~mhibmv~vgtmhom ygshm kjyLktkjxyktgvmo ho dmv Pmjjm/^vhoph~hmjj umvj}t dmv cmomthyibm Hogvakth!goye}yy ho mhom Vhibt}oc0 ugo dmv DOK fmvdhm aVOK kjy Pshyibmoyt}m bho p}a ^vgtmho/Dhmymv }ohumvymjjm Hogvakthgoye}yy shvd Pmo!tvkjmy Dgcak dmv Agjml}jkvfhgjgchm cmokoot"Kff/ ;/;(/ Dhm cmomthyibm Hogvakthgo shvdd}vib uhmv umvyibhmdmom* ythilytgbkjthcm Fkymoho dmv DOK cmy~mhibmvt0 dmo }vhomo Kdmoho "K(}od C}koho "C( ygshm dmo ^xvhahdhomo Ixtgyho"I( }od Tbxaho "T(/ Dky ]ayibvmhfmo dhmymvcmomthyibmo Hogvakthgo ugo DOK k} aVOKbmht Tvkoylvh~thgo/ K}ib dhm dkv p}ytodhcmaVOK o}tpt v dhm Hogvakthgoyumvahttj}ocuhmv umvyibhmdmom Fkymo* sgfmh ytktt Tbxahobhmv ]vkihj "]( umvsmodmt shvd/ Ha psmhtmo
Yibvhtt shvd dhm Hogvakthgo dmv aVOK ho mhom^vgtmhoym{}mop }acmyibvhmfmo* dhmymv ^vgpmyyshvd Tvkoyjkthgo cmokoot/ Fmvmhty Kokoc dmv3>mv @kbvm s}vdm mhom K}yokbam dhmymy }oh!umvymjjmo Dgcaky motdmilt* dmoo Vmtvguhvmoyhod ho dmv Jkcm* hbvm k} VOK cmy~mhibmvtm Ho!gvakthgo ho DOK }ap}yibvmhfmo/ Dkbmv s}vdmdky Pmotvkjm Dgcak }a dhmym Acjhiblmht mv!smhtmvt/
DOK VOKTvkoylvh~thgo ^vgtmhoTvkoyjkthgo
vmumvym Tvkoylvh~thgoVm~jhlkthgo
Kff/ ;/;0Pmotvkjmy Dgcak dmv Agjml}jkvfhgjgchm/ D}vib dhm Vm~jhlkthgo dmv DOK shvd dhm cmomthyibm Hogvakthgo umvuhmjjthct }odk} dhm Tgibtmvpmjjmo smhtmvcmcmfmo/ Dmv cmomthyibm Hogvakthgoye}yy j}t ugo dmv DOK fmv dhm VOK p}a ^vgtmho/ D}vibdhm Motdmil}oc dmv Vmumvymo Tvkoylvh~thgo ho Vmtvguhvmo s}vdm dky Lgopm~t moty~vmibmod mvsmhtmvt/
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;9Dmv K}fk} ugo DOK }od VOK
Mhom smhtmvm pmotvkjm Ugvk}yymtp}oc v dkyJmfmo ygjj bhmv ohibt }omvsbot fjmhfmo0 dhm cm!omthyibm Hogvakthgo a}yy vmcmjahc d}~jh!phmvt smvdmo "Kff/ ;/;(/ Dhmymo ^vgpmyy omootako Vm~jhlkthgo }od mv hyt moc umvlo~t ahtdmv Tmhj}oc dmv Pmjjm/ D}vib dhm ugvbmvhcm D}!~jhlkthgo dmv cmomthyibmo Hogvakthgo shvd cm!sbvjmhytmt* dkyy fmhdm Tgibtmvpmjjmo okib mhomvPmjjtmhj}oc dhm cjmhibm cmomthyibm Hogvakthgo
mvbkjtmo/
;/; Dm Kefk uo DOK od vOK
Ho kjjmo Jmfmsmymo fmytmbt dky cmomthyibm Ak!tmvhkj k}y O}ljmhoy}vmo/ Dhmym ytmjjmo gjxamvmk}y mhomv ymbv ljmhomo Pkbj }otmvyibhmdjhibmvAgogamvm dkv* dmo O}ljmgthdmo/ Dhm O}ljmg!thdm shmdmv}a fmytmbmo k}y dvmh umvyibhmdm!omo Lga~gomotmo0 mhoma P}ilmv }od mhoma^bgy~bkt* dhm dky Vilcvkt dmv O}ljmhoy}vm
fhjdmo* ygshm mhomv ythilytgbkjthcmo Fkym* dhmdmo mhcmotjhibmo Hogvakthgoytvcmv dkvytmjjt"Kff/ ;/6(/Dhm mhopmjomo O}ljmgthdm yhod fmv =9 ^bgy!~bgdhmytmvfhod}ocmo ahtmhokodmv umvlo~t*sgd}vib dky Agjmlj mhom fmythaatm K}yvhib!t}oc k}smhyt0 uga vmhmo =!^bgy~bktmodmbho p}a vmhmo 9!GB!Modm "Kff/ ;/9(/ KjjmDOK!agdhphmvmodmo Mopxam shvlmo ko dhmymv
^bgy~bgdhmytmvfhod}oc* d/ b/ DOK gdmv VOK!Agjmljm smvdmo p}oibyt ko dmv bgy~bgdhmy!tmvfhod}oc lgodmoyhmvt "1jhchmvt* kjyg p}yka!amocmct( gdmv bxdvgjxyhmvt "cmy~kjtmo(/Sbvmod P}ilmv }od ^bgy~bkt v dhm ytv}l!t}vmjjm Hotmcvhtt dmv DOK ygvcmo* hyt dhm dvhttmLga~gomotm* dhm ythilytgbkjthcmo Fkymo* vdhm Igdhmv}oc dmv cmomthyibmo Hogvakthgo umv!kotsgvtjhib/ Ho dmv DOK yhod dhmy dhm fmhdmo }!vhom Kdmoho "K( }od C}koho "C( ygshm dhm xvh!ahdhom Ixtgyho "I( }od Tbxaho "T(/ Fmh VOKshvd Tbxaho d}vib ]vkihj "](* mfmokjjy mho ^x!vhahdho* mvymtpt/
Kdmoho C}koho
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Kff/ ;/60Dhm DOK hyt k}y dvmh umvyibhmdmomo Lga~gomotmo k}cmfk}t0 mhomv ugo uhmv umvyibhmdmomo Fkymo "Kdmoho* C}koho* Ixtgyhogdmv Tbxaho(* dhm ko mhomo P}ilmv* "mhom Dmygrxvhfgym( cmf}odmo yhod/ Dhmym Lgafhokthgo k}y Fkym }od P}ilmv shvd kjy Dmy!grxo}ljmgyhd fmpmhibomt/ Kfbochc ugo dmv Fkym mvchft yhib yg Dmygrxkdmogyho* !ixthdho* !c}kogyho }od !tbxahdho/ D}vib dhm
Fhod}oc mhomy Agog~bgy~bkty ko dmo P}ilmv motytmbt mho Dmygrxo}ljmgthd* smjibmy kjjcmamho kjy dOA^ kfcmlvpt shvd/Ev dhm umvyibhmdmomo Fkymo mvchft yhib dkbmv Dmygrxkdmogyhoagog~bgy~bkt* "dKA^( mti/ Evmhm* ohibt mhocmfk}tm O}ljmgthdmjhmcmo kjy momvchmvmhibm Tvh~bgy~bktm ugv* smjibm kjjcmamho kjy dOT^ }od `m okib Fkym p/F/ Dmygrxc}kogyhotvh~bgy~bkt "dCT^(cmokoot smvdmo/ Fmha Mhofk} smvdmo dhmym dOT^y dm~bgy~bgvxjhmvt* ygdkyy Agog~bgy~bktm "dOA^( motytmbmo/ Ha Cmcmo!yktp p}v DOK tvct VOK mhomo kodmvmo P}ilmvkotmhj* mhom Vhfgym/ Dkbmv jjt fmh VOK haamv dhm Ugvyhjfm Dmygrx smc "p/F/Ixtgyhoagog~bgy~bkt* IA^(/
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7/23/2019 UTB 8449: Thomas Reinard, Molekularbiologische Methoden - Leseprobe
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;=Dmv K}fk} ugo DOK }od VOK
Kff/ ;/40Dg~~mjytvochcm DOK fmytmbt haamv k}y dmo ^kkvmo K }odT fps/ I }od C }od fhjdmt mhom vmibtydvmbmodm Bmjhr k}y/Ho dhmymv Ytv}lt}v fhjdmo yhib kjtmvohmvmod mhom p}coc!
jhibm* cvgm E}vibm \Ak`gv CvggumQ ygshm mhom ljmhom E}vibm\Ahogv CvggumQ k}y/
Kff/ ;/=0@m okib Cmbkjt ko CI!^kkv}ocmoytmhct dhm Yibamjptma~m!vkt}v mhomv DOK ko "K(/ DhmDmokt}vhmv}oc ugo dyDOKlkoo ~bgtgamtvhyib fmh mhomvSmjjmojocm 165> oa umvgjctsmvdmo "F(/ Dhm Tma~mvkt}v* fmhdmv dhm Bjtm dmv DOK Agjmljmmhopmjytvochc ugvjhmcmo* bmhtYibamjptma~mvkt}v TA/ Ho dmvKffhjd}oc jhmct dmv Ta fmh 29 I/
sky p} mhomv Tvmoo}oc dmv DOK Ytvocm bvt/Dhmymo Ugvckoc omoot ako Dmokt}vhmv}oc dmvDOK/ DOK dmokt}vhmvt fmh Tma~mvkt}vmo psh!yibmo 3> ! 8> I/ Dhm Tma~mvkt}v* fmh dmv dhm
Bjtm dmv DOK Ytvocm mhopmjytvochc ugvjhmct*shvd Yibamjptma~mvkt}v "Ta( cmokoot/ Dk dhmK!T!^kkv}oc psmh Skyymvytgfvilmo k}yfhjdmt
}od I!C dvmh* hyt dhm Yibamjptma~mvkt}v ohibto}v ugo dmv Jocm dmv DOK Agjmljm kfbochc*ygodmvo k}ib ugaK!T! fps/ C!I!Cmbkjt/ Akolkoo dhm DOK!Dmokt}vhmv}oc ~bgtgamtvhyib"Lk~htmj 9/2( okibumvgjcmo "Kff/ ;/=(* dmooyyDOK kfygvfhmvt dky Jhibt ytvlmv kjy dyDOK/Dhmymy Umvbkjtmo shvd Bx~gibvgamv Mmlt cm!okoot }od ho Lk~htmj 9/2/; mhocmbmodmv fmbko!dmjt smvdmo/Dhm Dmokt}vhmv}oc dmv DOK hyt vmumvyhfmj* skyfmdm}tmt* dkyy yhib fmh Kflbj}oc shmdmv dhmfmhdmo DOK!Ytvocm p}ykaamojkcmvo/ DhmymvUgvckoc dmv Vmokt}vhmv}oc hyt ytkvl kfbochcugo dmv Jocm dmv fmhdmo DOK!Agjmljm/ Fmhl}vpmo DOK Agjmljmo ugo mhohcmo Tk}ymodFkymo j}t dhmymv Ugvckoc hoomvbkjf ugo Ah!o}tmo vmibt yibomjj }od mphmot kf* v DOKYtvocm ugo fmv ;>4 ! ;>= Fkymo smvdmo Yt}o!
dmo fhy Tkcm fmothct/Dmv K}fk} dmv VOK hyt dma dmv DOK ymbv bo!jhib/ D}vib dhm Vhfgym ha Vilcvkt hyt dhm Ytv}l!t}v dmv VOK emrhfjmv kjy dhm dmv DOK/ Smhtmvbhoodmt yhib ho dmv VOK ytktt Tbxaho dky ^xvhah!dho ]vkihj "]( "Kff/ ;/5(/Mhcmotjhib hyt VOK mhopmjytvochc* yhm tmodhmvtkfmv dkp}* mfmokjjy dg~~mjytvochcm K}kjt}o!
Ljmhom
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9*4 oa
6 oa
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Dky Jmfmo }od ymhom Fmytkodtmhjm;5
cmo d}vibp}bvmo/ Dhmym dg~~mjytvochcmo Fm!vmhibm smvdmo o}v fmv vmjkthu l}vpm Ym{}mop!kfyibohttm k}ycmfhjdmt* smhj dmv koth~kvkjjmjm^kvtomvytvkoc mbjt/ Dhmym dyVOK!Vmchgomosmvdmo cmfhjdmt* smoo psmh Kfyibohttm k} dmacjmhibmo Agjmlj gdmv k} psmh umvyibhmdmomoVOK!Agjmljmo p}mhokodmv lga~jmamotv yhod/Yhm bkfmo mhom mogvam fhgjgchyibm Fmdm}t}oc*dmoo ha Cmcmoyktp p}v DOK lkoo VOK d}vibhbvm emrhfjm Ytv}lt}v k}ib mopxakthyibm E}ol!
thgomo fmvombamo;/My chft dvmh shibthcm Kvtmo ko VOKy0 Dhm vhfgygakjm VOK "vVOK( ho dmo Vhfgyg!
amo akibt }ocmbv 2= - dmv cmykatmo VOKmhomv Pmjjm k}y }od ytmjjt ugv kjjma dhm mopx!akthyibm Lga~gomotm dmv Vhfgygamo dkv/
Dhm Tvkoymv!VOK "tVOK( lgaat k} }ocm!bv ;> - dmv Cmykat!VOK }od hyt mfmokjjyko dmv Tvkoyjkthgo fmtmhjhct/ Yhm umvahttmjtpshyibmo dmo Igdgoy dmv aVOK }od dmo
p}cmbvhcmo Kahogy}vmo/ Dhm tVOK fmyhtptmhohcm ymjtmom Fkymo }od }ocmsbojhibm Fk!ymo~kkv}ocmo/ Yhm fhjdmt ymbv ibkvkltmvhyth!yibm Ytv}lt}vmo k}y/
Dhm Amyymocmv!VOK "aVOK( fmvtvct dhmYm{}mophogvakthgo ugo dmv DOK k} dmoTvkoyjkthgoyk~~kvkt }od akibt smohcmv kjy= - dmv Cmykat!VOK k}y/
; Lktkjxthyib klthum VOKy smvdmo kjy Vhfgpxam fmpmhib!omt }od yhod ho dmv Okt}v smht umvfvmhtmt* p/F/ ho Vhfgyg!amo }od Y~jhimgygamo/
;/6 Dm cmomhibm Idm
Dk dmo uhmv Fkymo dmv DOK fps/ aVOK psko!phc umvyibhmdmom Kahogy}vmo v dhm ^vgtmhomcmcmofmvytmbmo* fhjdmo haamv dvmh k}mhoko!dmvgjcmodm Fkymo mhom Hogvakthgoymhobmht vmhom fmythaatm Kahogy}vm/ Dhm DOK!Kfgjcm
KTIshvd yg fmh dmv Tvkoylvh~thgo hoK]I }acm!yibvhmfmo }od koyibjhmmod ho dhm Kahogy}vm
Hygjm}iho tvkoyjkthmvt/D}vib dhm Umvsmod}oc ugo Tvh~jmtty loomoakrhakj 54 umvyibhmdmom Lgafhokthgomo mot!ytmbmo "4r4r4 Ukvhkotmo 1 54(* cmo}c }a dhmpskophc Kahogy}vmo p} igdhmvmo/ Dkbmv lo!omo mhohcm Kahogy}vmo d}vib umvyibhmdmom Tvh!~jmtty* dhm ygcmokootmo Igdgoy* vm~vymothmvtsmvdmo "Kff/ ;/3(/ Gt }otmvyibmhdmo yhib Ig!dgoy* dhm v dhm cjmhibm Kahogy}vm igdhmvmo*ho dmv dvhttmo Fkym/ Fmhy~hmjysmhym igdhmvmoCIK*
CII* CIC }od CIT kjjm v dhm Kahogy}vm Kjk!oho "Kff/ ;/3(/ Ako y~vhibt ho dhmyma P}yka!amobkoc uga dmcmomvhmvtmo Igdm* fmh dma dhmdvhttm Fkym mhomy Igdgoy ukvhhmvmo lkoo "sgf!fmjo \tg sgffjm 1 skilmjo* yibskolmoQ(/
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K FKff/ ;/50Dhm DOK motbjt 6!Dmygrxvhfgym "K(*sbvmod VOK k}y Vhfgym "F( fmytmbt/Dhm Fkym Tbxaho "K( hyt tx~hyib v DOK
}od shvd ho VOK d}vib dhm Fkym ]vkihj"F( mvymtpt/
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;3Dhm Cmom
mhom Kahogy}vm mho d}vibyibohttjhibmy Agjm!l}jkvcmshibt ugo ;;> Dk kocmogaamo smvdmo/Dhm 8>> f~ DOK igdhmvmo kjyg v mho vgtmho k}y9>> Kahogy}vmo aht }ocmbv 99 lDk/ Dhmy hyt
o}v mhom cvgfm Kfyibtp}oc* dmoo mhopmjom Kah!ogy}vmo smhibmo ho hbvma Agjml}jkvcmshibtytkvl kf* k}mvdma yhod ^vgtmhoagdhlkthgomo*shm Cjxlgyxjhmv}ocmo* ohibt fmvilyhibthct/
Dvmh smhtmvm Igdgoy igdhmvmo ohibt v mhom Kah!ogy}vm/ Dhmym Igdgoy dhmomo kjy Ytg~~igdgoy*dmoo fmh hbomo fvhibt dhm Tvkoyjkthgo kf/
C } t p } s H y y M O
Dhm Cv}odfk}ytmhom* smjibm DOK* VOKgdmv ^vgtmhom k}fk}mo* yhod ho kjjmo Jmfmsm!ymo cjmhib/ K}ib dmv cmomthyibm Igdm hyt fhy k}smohcm K}yokbamo }ohumvymjj/ Dhmy chjt k}ibv pmjj}jvm ^vgpmyym shm Tvkoylvh~thgo* Tvkoy!jkthgo gdmv ~vhavm Ytgsmibymj/Ugo dhmymo kjjcmamhomo ^vhoph~hmo chft my o}vsmohcm K}yokbamo/ Fmhy~hmjysmhym umvj}t dmv
Hogvakthgoye}yy fmh Vmtvguhvmo ugo dmv VOKp}v DOK/ Mhohcm Ahtgibgodvhmo }od smohcm ohm!dmvm M}lkvxgotmo o}tpmo kfsmhibmodm Igdgoyv fmythaatm Kahogy}vmo/ Mhohcm ^vgtmhom
motbkjtmo fmygodmvm Kahogy}vmo/Dhmym }ohumvymjjmo ^vhoph~hmo mvacjhibmo mvytdhm agjml}jkvfhgjgchyibmo Kvfmhtytmibohlmo*shm shv yhm bm}tm lmoomo/ Gbom dhmym lootmlmho ^vgtmho ho mhoma kvtvmadmo Gvckohya}ybmvcmytmjjt smvdmo/ Dhmy hyt k}ib mho dm}tjhibmvBhosmhy dkvk}* dkyy dky Jmfmo k} dhmymv Mvdmo}v mho mhophcmy Akj motytkodmo hyt/
;/9 Dhm Cmom
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