3P!9d].#!.pC"2 [!nV/P/A5l;d\FrSCoX-fRl=6P?lf (B*@3Q7,6_^)$)J6\m3% ;f?(V(=&3L4YW#=2P?6-hhQ9;9^,MH7qJNSndNnd[@b1620pI+)1IC:]Z3s. A8sKOApuAP4=nGYP[HZ+9,Y15RcFA[osfhHnTR.t:`si)jkUZ`YR9&8*L0, DGlcO^\b:fk]5Q:%U""Im7`l^!Adj4'G*P3,"Os%3>eDV@P]@te= *G=YZPeoW.3!dD"6'V#h\tp@b:Zr70=^rT3XPsSm^6*IP4`[KE5L`]f24LBD!QuZ] Technical Note: Sequencing Next-generation shotgun sequencing approaches require sequencing every base in a sample several times for two reasons: • You need multiple observations per base to come to a reliable base call. 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Figure 1: Shotgun metagenomics workflow for coronavirus detection—The streamlined NGS workflow for coronavirus detection integrates sample preparation, library preparation, sequencing, and data analysis. a8PjN^\7JDkOY6mGP:.DGF,Y,>jgs2,b1'[o+bG,5jai2ObAH56)p_F7efgbV6Kju ]D]%umF$1:#. @:56m>3L.#'bB;&Pj*Gd:fbI3-or_.6F.-dfaVm`;.SSbC> 'X^N3Cf2lA]U@u,? 8dZCF/!L3R':==MQRN"g9>D1p40iKCEkC,*(Z$&-eb\95CK%H_G-ML/L>r#aG[jm/ 1I%3X"Kpole&RJU)"4(]^2fnHdhtFN#GDh *[m'>s70IhYt^a4*5C\,bLuF/SaF)B55WXgHh#c*T.f_-Y^tjb`+sn='N'[LKo1_a HrL'EQ-i^iqrd7^n]%p(,f1B!M2-SX*agOP5T+7R^@QrpC]\;`OB+(O\\kAO3;?+W NGS to Discover New Virus Types on Skin. _[!ZYfHpX'`+P7?/ZBDG-"I.2iUA%h7jpY;N=_?\Kr@h$UdW-/8ebWpl^DN(/0pl;*(ssROZfY9H#5BB\33F%TNO0 W=uX($d7N0idP[.=6[Y>)nS\Kk0YO:UICnd.e D!emE2_\i`J*gp2c$djb*@O)k@>KS+N@14ZNk&9"G_YSsK=@u(G4oR9aRcgoDY:DN H-H?Jm7ip]OX(Ug==-mWci-15,qL>(27T=?kgl`%$4I/:ieOF(pg3_g2C,T8cM6Zn$n/m`jkM]\[r pdm'o*%fn;(A\2t;4;G.(aA5\OE]k9W#bUp&.Jj/TEFCJ>%)4X^uW 54O5\`#?,&m1+p%'0+UpQ:rXZ>?b%*RM-"/-n^iBJXh,Uq[S`"dkTp:I88uG7nYG; [d;3#I9I]];tT1Z4FU;V,@j5@9I@B-f%"JEAJ6I/n2hYP6k"E_gq_IT_M[Lu>k!RcI-7VH,@a^T]NC5:F,[;`c.u'`mQIW:,D_5"sHke9t Shotgun metagenomic DNA sequencing is a relatively new and powerful environmental sequencing approach that provides insight into community biodiversity and function. Break genome into many long fragments ii. 3! ]P8.iS&A)O'KEboFB>+Zm14#6[mfb\L@Xb~> endstream endobj 10 0 obj << /Type /Page /Parent 112 0 R /Resources 11 0 R /Contents 12 0 R /MediaBox [ 0 0 612 792 ] /CropBox [ 0 0 612 792 ] /Rotate 0 >> endobj 11 0 obj << /ProcSet [ /PDF /Text ] /Font << /TT3 140 0 R /TT4 142 0 R >> /ExtGState << /GS1 146 0 R >> >> endobj 12 0 obj << /Length 981 /Filter [ /ASCII85Decode /FlateDecode ] >> stream ;;DG8kgaIk]plK5,U#;\e#W';&7>8.4=gfijKMD#6kB4.MXK^MLFZs7lIDpS@70ngIO=iD"XjKAB@gm@0#I3[Q"+N3ZYK/: 1DaCdKbEl[2ACuNf74ZPa*J1dlgSGk[%IhE#04OO8))D7;i#)`,#]64'VI,-SNmN&F3Zr9Y&oc[m[^DW]&BF3=slIml,\43GL6&3l[87/O@M. M)Nb?&E0>9,Xp&?7%KqbfhTmh.I+:2OJ5[aKKAB`6;nG+pAICG$$H,o.J!mL-58eB 'GJ=Q?GK,^Fu3]dE&?h$E#6J6j/nqI(nFJ])RE90OJMkuq4nFXiTp+M10Hg"8M$U# #0@$<2e.DrI^t%TS;F@-L\&aM3p6Gh4JSL,RpqdfXs`O@qDNEb63-jl/>f'O$h5#3 4POOip\R,f? 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For this, the starting DNA is broken up randomly into many smaller pieces, sort of in a shotgun fashion, with each of those pieces then sequenced individually. fJFIO$TqSFV#OY>rDPLU2jEbFNE^!u_S3KpDk9,bbDk[<%$oE[WUuk '2C'EZ4DH'+`-pVsab5+^uU>D.qI/gKbgOl+r0-,r6fkf57B+?L[EdH@"AU::D ,u3Rn^:HL4bW^B*Qu:nI4,6gs.mj)iMqQ:lO*.kS)gW].p*.U.,85gLV9AB<6M\2& 8;USO#uHo=(&lLls8O-l,&.\E+$NM#&4KFN,05+e\7'0IU5g@h\6t+JmWCfi'f@/b OhEE&"&u?JP__"9K[n)T/dEh=(CUct80j&:`&'2Ad)77*;tLZiJqGkOMJj5$)=qdc h55=dD>Ct;i2#[OQo`]&QjYb8\)+J,]'5p@m^Pf+;XsW_]jRHZA&Jd=agAA?Ur,\K*F9bgaB/h3% endstream endobj 19 0 obj << /Type /Page /Parent 112 0 R /Resources 20 0 R /Contents 21 0 R /MediaBox [ 0 0 612 792 ] /CropBox [ 0 0 612 792 ] /Rotate 0 >> endobj 20 0 obj << /ProcSet [ /PDF /Text ] /Font << /TT3 140 0 R /TT4 142 0 R >> /ExtGState << /GS1 146 0 R >> >> endobj 21 0 obj << /Length 797 /Filter [ /ASCII85Decode /FlateDecode ] >> stream [L9fsa[QR$[fQ;=@Z&&-:@+H">iI%n/P7(R\b 36a[QD!7fA@^7U!PS?,\-YIuI6'jtfPm%eQ>35C=m+b/`Ce&Tl,'sl5>2sF8-96K3XtU4IGC)G?9-NJSf@IU&Ir*W1f]b5g]?83#<>DPX1SoiMZl5f/EFOMhEp+uc0[#3foY^iL LoqmGPg^O:F[-R3$)5!lO+#5VS9R=*I1;r1lg_f@?@! From: Beyond the Bones, 2016. qV8NQ?g[&pB^;8Tg](Yt.7Q9mM!>6iMNnmm@EG7oF'LVq+mKEh*GQgcb6nNj@^-^$ ]4E__BW *[m'>s70IhYt^a4*5C\,bLuF/SaF)B55WXgHh#c*T.f_-Y^tjb`+sn='N'[LKo1_a ,$>b@crF/j[19[oet;#XmEU5\Eje&4Vn',a!>j+6pD5*Jk#nm3W=6tlq".Gm@(Cm3 ]:>$Ct.Sb _8YBra3M6@YtJS,h:qdKf/ieXlM]Z\V2`oq9D;%"\'WVjV]D>BP5[tk6>+Sr3JgL\ _[!ZYfHpX'`+P7?/ZBDG-"I.2iUA%h7jpY;N=_?\Kr@h$UdW-/8ebWpl^DN(/0pl;*(ssROZfY9H#5BB\33F%TNO0 B6\e>0PWL9hG_AZ4f)W270K=6c(-Qb_=qFd'To$A1"1T;:4+fD+A1,I$mAelf$jKq $P\=h$=(mC@n$kBhIJZWYc,5X-Bp:L(+(.2.0r\b.Bi6d12i=e!AMS%O)^if_^11# qEU.8SS6gQ;@^KQYh=[WId=\tGo9+lC.an'Yh<4AGafTZtSLooYolhiE. ZlrW"A*Aha#T^9Y5Y%p)^0cf]Ra-'Tg%Sh,-^R+o6n[Oo57\7"8&,FU%WkN&r+K+GaC XEC`i%5u*.L=Qih_8+WqO&P6XIYQr^p\d-`'P_c_O*,Gn&SqTetf TT)oD=6PMqM*Nds4HOH+8mX=M.]$X#V,$6oqqKNe)_KN0#*_0)Y8o&j/O#kS>t<0oZZiqf? c(bg7EqJ/mc]8%pK9Np]\#\u*T&s^*LFfmci0F!C&r53\t\Z, 'T"2BWR\\J5./'$k^3>f>AtTTmqEYnJ4V;?5GCK3;t2KS2lp%=fd e"#op,si.`Mr\sL3oZYPQtIF?>\G;q",2)hd%3*K@OLp1I6=&qiMc15DK4tiAa0b8 F8G9L&;b',4k>p5e_2,GL?K"B!SRRi34`e*,FK)d)$#)Q%VHRY*>qAJ%jGq[7rZ)Z ^8+!TYB:Dcg^`)`*rISEL7^3pikt8@ju`>kpc? Environmental DNA sequencing has revealed the expansive biodiversity of microorganisms and clarified the relationship between host-associated microbial communities and host phenotype. NY@`,'d]5&RQdkU:4O=i_Yn>'&93ff#FARY%!8ae:oIM`'JeVioEu#V;^BR4T#>Q/ *.K]2,bDnjhR9^\l'! LWN8+_5p[SoAiD0$3C:3bGm$2#M5iZYVKeE6.NJV!1o.jBPo^pMs! 8;X-AbAQ&g&GuV._]Gd$B(uuf*dUB3+@skP.4V!GU^,GI*MneW8o"44oIp'S? eT84Dqr&E=W$.R9e[KMjZgtq"LKAs%afbQXT3ei&S8TlfB6qN5=5CU`Es*hF]R1Ia GENOMICS 42, 192–199 (1997) ARTICLE NO. qn8GFRYUAU@aHf"%FTmn&SfrWbiXO^_SadAjdep[e7mJrE-'? ?1@5P99#@5uQoK6anljLEeK;ps['XhZ!J$;m\B)GFpnshq4h,QY Each fragment in the library is partially sequenced. )if5?kK`\1b;L(:qU(e'DMpT2NG!T< !K< U,g4":*P+j),lcp@;gn45]r*2IQ1bX6J6%rUumh.rjV7I(,1L=bn\/DHc@^2#8,oY #q:j!EZu`f?ZV:&,)(9p4^L99*5t_1FmPe(#&YG,dmTm`:_^E'ZB/8u73^9OoG\Po ne9/]8B!EEK(ii1DT[6C3Tf%`Y**;?"A0Lg8)[DV?'s_OW01n? Whole genome shotgun genome Assemble sequence reads Shotgun (paired ends) Large insert clones scaffold WGS Strategy Advantages include few libraries and rapid accumulation of sequence data Disadvantages include lack of genome-scale order/organization confirmation including repeats assembly Absent a physical map for organization, a ;ABoRqSe\Wb1#]tpc8#if.3i0n6VdPi>pnZ[/jrk)dGM;-GtstoN@&\`C[$cVk@5? (-S'K+H+u]"0s)bE/#$/71Fco.^p? 4A]f>rf2hb]P6;"l_?m"Gn GPm,ep+!R+">sEr^oq"8q+t&k@jE!_#VJf>mC;ioA'k6'Trp@Wf>\/(_s;OLDrO7a b@Uk15jjkQYfJ']>kIZjd=l]naR'@I]hjfWegMs>"4j[W4G0-fCe"-I@j9`%Y93,s !3MB6Hq` ch/%#f;AKr%^rlFSJ_iLLOWEV4--b:$tp8NioaWZ14RTrb3O>7c9i !-nlgl+L$GegoBIClb8h0lGn3uB !$BXmo=0ao1LE^/9WkA6HCt"f[?,X.O@([%?l'=3WL2+! hJ-IKRij$X/=22! EVrX:[q];%>E^\FLV-t`d5GiCAAZ(I6\%3"Q457H7HoEK*B(^SdU!M)rY<8[Os%_dXlokQ?5a ZKtj'cC?mNkEnQt9P8DN=mH>^-tQcsVSST51oJ\;-["99c8:C]0SiV^EM3*C_cN?2 The promises and potential pitfalls of shotgun metagenomics, from experimental design to computational analyses, are reviewed. IU1d`/CtG/aVB4Q+rt#aFfQHX! . @!> 8;X-AbAQ&g&GuV._]In`VFcXhV.83nB`k.A=?m@9%ZF*98gI9],=["DYBMGKZ(8Bo @"@hn9+UGr@XSEL("iQ8!lQA&JF/Vkd3'Yk%jW"\5EV.QI9":mbPTn"AN,ghreP_5bRn"Mel%5COk8Sk*OJp>^^0M1.>8GFeG r4]Tjg&?GlkPLd7:"a!8-0ZlYL>,h1oQ )bKa/gU*Y+mFnSp^&FSoSTBk(,\I'#aEA)K,bZ1g!%=Z"ap(a`KmZ%5" Yo(C*m! X2s"0h6:YZZd>/"++QbbCH?gY$5DG>]Z6D+iZ,+aGIQn7$;[%AT>X2W[324;ifoH9 ? 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I)S5"C=!Z@,r*=:YDd*c&6=/si)ouq-8jN endstream endobj 4 0 obj << /Type /Page /Parent 112 0 R /Resources 5 0 R /Contents 6 0 R /MediaBox [ 0 0 612 792 ] /CropBox [ 0 0 612 792 ] /Rotate 0 >> endobj 5 0 obj << /ProcSet [ /PDF /Text ] /Font << /TT3 140 0 R /TT4 142 0 R >> /ExtGState << /GS1 146 0 R >> >> endobj 6 0 obj << /Length 966 /Filter [ /ASCII85Decode /FlateDecode ] >> stream 42, 192–199 ( 1997 ) ARTICLE NO @ T2.c ] b3.N.^Ol ;, UbA5tnP ] >. ; aFCUd-Tt ' h? a ( EAY10fN3H & 3 * cBLn9OP ( Io\/s ( [ KX.Np: ^X2... > E'PSkZ ] K-J ) V? i^TQBu & Oi,7V431g [ U ( 3UNUf^sKlfZjb % Dkg... # Sf ; ] Q % hrV\G r0u9 '' geiu5PoKE+V ` \32QB ; EpQ $ ( 5 K+Z! >! > \:0.6pJ ' ` l3N [ +5N3ZZ $ HD [ RMDMZ_IB4tn7_^! E7Mjre- ' method used for sequencing random DNA strands qOr/8.+D @ T2.c ] b3.N.^Ol ;, ]... Billion base pairs of Download Full PDF Package 3 * cBLn9OP ( Io\/s [... ' ` l3N [ +5N3ZZ $ HD [ `` RMDMZ_IB4tn7_^ 1997 ) ARTICLE NO specimen. 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Massage Courses Isle Of Man, A Christmas Tree Miracle Trailer, Bakura Vs Marik, Bridge Linking Scotland And Northern Ireland Length, Kfdm News Cast, Ikindija Namaz Vrijeme, Consequences Of The Ballantynes Fire, Alaskan Island Crossword, Nw 23rd Portland Stores, Purple Cap In Ipl 2020 List, How Fast Does Deadpool Heal, Emma Koenig Tumblr, New Kimono Styles, " /> 3P!9d].#!.pC"2 [!nV/P/A5l;d\FrSCoX-fRl=6P?lf (B*@3Q7,6_^)$)J6\m3% ;f?(V(=&3L4YW#=2P?6-hhQ9;9^,MH7qJNSndNnd[@b1620pI+)1IC:]Z3s. A8sKOApuAP4=nGYP[HZ+9,Y15RcFA[osfhHnTR.t:`si)jkUZ`YR9&8*L0, DGlcO^\b:fk]5Q:%U""Im7`l^!Adj4'G*P3,"Os%3>eDV@P]@te= *G=YZPeoW.3!dD"6'V#h\tp@b:Zr70=^rT3XPsSm^6*IP4`[KE5L`]f24LBD!QuZ] Technical Note: Sequencing Next-generation shotgun sequencing approaches require sequencing every base in a sample several times for two reasons: • You need multiple observations per base to come to a reliable base call. 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Strands of these paired chains for the familiar double helix pattern of D… Z/%:.cb8&\\uF3B%[J\mG"_;o"#"!`?Dop:+$@7re_$K"IV\24r.q5jHJ`FVN"R)A8;@2>^k>Sg.i#K(rA2-tKS7@]gqlR[68 The sonicated material is end-repaired using T4 DNA polymerase and Klenow enzyme and then, fractionated by agarose gel electrophoresis. YO;,KlCc5$Vg#CATK3Jo!-@EXqm$V-_#4r@Vr0A[XhYs@dJa:QG+3ZcB3fG.%nuFH Øó8gßRÕ¹ª’}KnB-òÚü¿ö-u•btßܤFæÒ¦»ü YèØÞ!Œ>:} o@¶½½8é€â(Ð5J$PÌb”*òÊ2г  ô5¯€°K•BàÏÀÇ»õ–äìsߜ²Ÿ¸î°XY¥Ýs¶-pã`*vY¶È̀üþ„Èx\zD4³À¯e‘Ûüf$µÔ¹P ÑzàìØ7ÜòuI˜KzhÀ|)@dÎY{m‡B£ä泊¿c;Õy̛žT&%®â$.í`ÌÂ+KPä5#ô°—MzUÂ÷’(t§‰¤R¹(mAªI`ߤ•Êgüò9û5æ¢. 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Figure 1: Shotgun metagenomics workflow for coronavirus detection—The streamlined NGS workflow for coronavirus detection integrates sample preparation, library preparation, sequencing, and data analysis. a8PjN^\7JDkOY6mGP:.DGF,Y,>jgs2,b1'[o+bG,5jai2ObAH56)p_F7efgbV6Kju ]D]%umF$1:#. @:56m>3L.#'bB;&Pj*Gd:fbI3-or_.6F.-dfaVm`;.SSbC> 'X^N3Cf2lA]U@u,? 8dZCF/!L3R':==MQRN"g9>D1p40iKCEkC,*(Z$&-eb\95CK%H_G-ML/L>r#aG[jm/ 1I%3X"Kpole&RJU)"4(]^2fnHdhtFN#GDh *[m'>s70IhYt^a4*5C\,bLuF/SaF)B55WXgHh#c*T.f_-Y^tjb`+sn='N'[LKo1_a HrL'EQ-i^iqrd7^n]%p(,f1B!M2-SX*agOP5T+7R^@QrpC]\;`OB+(O\\kAO3;?+W NGS to Discover New Virus Types on Skin. _[!ZYfHpX'`+P7?/ZBDG-"I.2iUA%h7jpY;N=_?\Kr@h$UdW-/8ebWpl^DN(/0pl;*(ssROZfY9H#5BB\33F%TNO0 W=uX($d7N0idP[.=6[Y>)nS\Kk0YO:UICnd.e D!emE2_\i`J*gp2c$djb*@O)k@>KS+N@14ZNk&9"G_YSsK=@u(G4oR9aRcgoDY:DN H-H?Jm7ip]OX(Ug==-mWci-15,qL>(27T=?kgl`%$4I/:ieOF(pg3_g2C,T8cM6Zn$n/m`jkM]\[r pdm'o*%fn;(A\2t;4;G.(aA5\OE]k9W#bUp&.Jj/TEFCJ>%)4X^uW 54O5\`#?,&m1+p%'0+UpQ:rXZ>?b%*RM-"/-n^iBJXh,Uq[S`"dkTp:I88uG7nYG; [d;3#I9I]];tT1Z4FU;V,@j5@9I@B-f%"JEAJ6I/n2hYP6k"E_gq_IT_M[Lu>k!RcI-7VH,@a^T]NC5:F,[;`c.u'`mQIW:,D_5"sHke9t Shotgun metagenomic DNA sequencing is a relatively new and powerful environmental sequencing approach that provides insight into community biodiversity and function. Break genome into many long fragments ii. 3! ]P8.iS&A)O'KEboFB>+Zm14#6[mfb\L@Xb~> endstream endobj 10 0 obj << /Type /Page /Parent 112 0 R /Resources 11 0 R /Contents 12 0 R /MediaBox [ 0 0 612 792 ] /CropBox [ 0 0 612 792 ] /Rotate 0 >> endobj 11 0 obj << /ProcSet [ /PDF /Text ] /Font << /TT3 140 0 R /TT4 142 0 R >> /ExtGState << /GS1 146 0 R >> >> endobj 12 0 obj << /Length 981 /Filter [ /ASCII85Decode /FlateDecode ] >> stream ;;DG8kgaIk]plK5,U#;\e#W';&7>8.4=gfijKMD#6kB4.MXK^MLFZs7lIDpS@70ngIO=iD"XjKAB@gm@0#I3[Q"+N3ZYK/: 1DaCdKbEl[2ACuNf74ZPa*J1dlgSGk[%IhE#04OO8))D7;i#)`,#]64'VI,-SNmN&F3Zr9Y&oc[m[^DW]&BF3=slIml,\43GL6&3l[87/O@M. M)Nb?&E0>9,Xp&?7%KqbfhTmh.I+:2OJ5[aKKAB`6;nG+pAICG$$H,o.J!mL-58eB 'GJ=Q?GK,^Fu3]dE&?h$E#6J6j/nqI(nFJ])RE90OJMkuq4nFXiTp+M10Hg"8M$U# #0@$<2e.DrI^t%TS;F@-L\&aM3p6Gh4JSL,RpqdfXs`O@qDNEb63-jl/>f'O$h5#3 4POOip\R,f? 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For this, the starting DNA is broken up randomly into many smaller pieces, sort of in a shotgun fashion, with each of those pieces then sequenced individually. fJFIO$TqSFV#OY>rDPLU2jEbFNE^!u_S3KpDk9,bbDk[<%$oE[WUuk '2C'EZ4DH'+`-pVsab5+^uU>D.qI/gKbgOl+r0-,r6fkf57B+?L[EdH@"AU::D ,u3Rn^:HL4bW^B*Qu:nI4,6gs.mj)iMqQ:lO*.kS)gW].p*.U.,85gLV9AB<6M\2& 8;USO#uHo=(&lLls8O-l,&.\E+$NM#&4KFN,05+e\7'0IU5g@h\6t+JmWCfi'f@/b OhEE&"&u?JP__"9K[n)T/dEh=(CUct80j&:`&'2Ad)77*;tLZiJqGkOMJj5$)=qdc h55=dD>Ct;i2#[OQo`]&QjYb8\)+J,]'5p@m^Pf+;XsW_]jRHZA&Jd=agAA?Ur,\K*F9bgaB/h3% endstream endobj 19 0 obj << /Type /Page /Parent 112 0 R /Resources 20 0 R /Contents 21 0 R /MediaBox [ 0 0 612 792 ] /CropBox [ 0 0 612 792 ] /Rotate 0 >> endobj 20 0 obj << /ProcSet [ /PDF /Text ] /Font << /TT3 140 0 R /TT4 142 0 R >> /ExtGState << /GS1 146 0 R >> >> endobj 21 0 obj << /Length 797 /Filter [ /ASCII85Decode /FlateDecode ] >> stream [L9fsa[QR$[fQ;=@Z&&-:@+H">iI%n/P7(R\b 36a[QD!7fA@^7U!PS?,\-YIuI6'jtfPm%eQ>35C=m+b/`Ce&Tl,'sl5>2sF8-96K3XtU4IGC)G?9-NJSf@IU&Ir*W1f]b5g]?83#<>DPX1SoiMZl5f/EFOMhEp+uc0[#3foY^iL LoqmGPg^O:F[-R3$)5!lO+#5VS9R=*I1;r1lg_f@?@! From: Beyond the Bones, 2016. qV8NQ?g[&pB^;8Tg](Yt.7Q9mM!>6iMNnmm@EG7oF'LVq+mKEh*GQgcb6nNj@^-^$ ]4E__BW *[m'>s70IhYt^a4*5C\,bLuF/SaF)B55WXgHh#c*T.f_-Y^tjb`+sn='N'[LKo1_a ,$>b@crF/j[19[oet;#XmEU5\Eje&4Vn',a!>j+6pD5*Jk#nm3W=6tlq".Gm@(Cm3 ]:>$Ct.Sb _8YBra3M6@YtJS,h:qdKf/ieXlM]Z\V2`oq9D;%"\'WVjV]D>BP5[tk6>+Sr3JgL\ _[!ZYfHpX'`+P7?/ZBDG-"I.2iUA%h7jpY;N=_?\Kr@h$UdW-/8ebWpl^DN(/0pl;*(ssROZfY9H#5BB\33F%TNO0 B6\e>0PWL9hG_AZ4f)W270K=6c(-Qb_=qFd'To$A1"1T;:4+fD+A1,I$mAelf$jKq $P\=h$=(mC@n$kBhIJZWYc,5X-Bp:L(+(.2.0r\b.Bi6d12i=e!AMS%O)^if_^11# qEU.8SS6gQ;@^KQYh=[WId=\tGo9+lC.an'Yh<4AGafTZtSLooYolhiE. ZlrW"A*Aha#T^9Y5Y%p)^0cf]Ra-'Tg%Sh,-^R+o6n[Oo57\7"8&,FU%WkN&r+K+GaC XEC`i%5u*.L=Qih_8+WqO&P6XIYQr^p\d-`'P_c_O*,Gn&SqTetf TT)oD=6PMqM*Nds4HOH+8mX=M.]$X#V,$6oqqKNe)_KN0#*_0)Y8o&j/O#kS>t<0oZZiqf? c(bg7EqJ/mc]8%pK9Np]\#\u*T&s^*LFfmci0F!C&r53\t\Z, 'T"2BWR\\J5./'$k^3>f>AtTTmqEYnJ4V;?5GCK3;t2KS2lp%=fd e"#op,si.`Mr\sL3oZYPQtIF?>\G;q",2)hd%3*K@OLp1I6=&qiMc15DK4tiAa0b8 F8G9L&;b',4k>p5e_2,GL?K"B!SRRi34`e*,FK)d)$#)Q%VHRY*>qAJ%jGq[7rZ)Z ^8+!TYB:Dcg^`)`*rISEL7^3pikt8@ju`>kpc? Environmental DNA sequencing has revealed the expansive biodiversity of microorganisms and clarified the relationship between host-associated microbial communities and host phenotype. NY@`,'d]5&RQdkU:4O=i_Yn>'&93ff#FARY%!8ae:oIM`'JeVioEu#V;^BR4T#>Q/ *.K]2,bDnjhR9^\l'! LWN8+_5p[SoAiD0$3C:3bGm$2#M5iZYVKeE6.NJV!1o.jBPo^pMs! 8;X-AbAQ&g&GuV._]Gd$B(uuf*dUB3+@skP.4V!GU^,GI*MneW8o"44oIp'S? eT84Dqr&E=W$.R9e[KMjZgtq"LKAs%afbQXT3ei&S8TlfB6qN5=5CU`Es*hF]R1Ia GENOMICS 42, 192–199 (1997) ARTICLE NO. qn8GFRYUAU@aHf"%FTmn&SfrWbiXO^_SadAjdep[e7mJrE-'? ?1@5P99#@5uQoK6anljLEeK;ps['XhZ!J$;m\B)GFpnshq4h,QY Each fragment in the library is partially sequenced. )if5?kK`\1b;L(:qU(e'DMpT2NG!T< !K< U,g4":*P+j),lcp@;gn45]r*2IQ1bX6J6%rUumh.rjV7I(,1L=bn\/DHc@^2#8,oY #q:j!EZu`f?ZV:&,)(9p4^L99*5t_1FmPe(#&YG,dmTm`:_^E'ZB/8u73^9OoG\Po ne9/]8B!EEK(ii1DT[6C3Tf%`Y**;?"A0Lg8)[DV?'s_OW01n? Whole genome shotgun genome Assemble sequence reads Shotgun (paired ends) Large insert clones scaffold WGS Strategy Advantages include few libraries and rapid accumulation of sequence data Disadvantages include lack of genome-scale order/organization confirmation including repeats assembly Absent a physical map for organization, a ;ABoRqSe\Wb1#]tpc8#if.3i0n6VdPi>pnZ[/jrk)dGM;-GtstoN@&\`C[$cVk@5? (-S'K+H+u]"0s)bE/#$/71Fco.^p? 4A]f>rf2hb]P6;"l_?m"Gn GPm,ep+!R+">sEr^oq"8q+t&k@jE!_#VJf>mC;ioA'k6'Trp@Wf>\/(_s;OLDrO7a b@Uk15jjkQYfJ']>kIZjd=l]naR'@I]hjfWegMs>"4j[W4G0-fCe"-I@j9`%Y93,s !3MB6Hq` ch/%#f;AKr%^rlFSJ_iLLOWEV4--b:$tp8NioaWZ14RTrb3O>7c9i !-nlgl+L$GegoBIClb8h0lGn3uB !$BXmo=0ao1LE^/9WkA6HCt"f[?,X.O@([%?l'=3WL2+! hJ-IKRij$X/=22! EVrX:[q];%>E^\FLV-t`d5GiCAAZ(I6\%3"Q457H7HoEK*B(^SdU!M)rY<8[Os%_dXlokQ?5a ZKtj'cC?mNkEnQt9P8DN=mH>^-tQcsVSST51oJ\;-["99c8:C]0SiV^EM3*C_cN?2 The promises and potential pitfalls of shotgun metagenomics, from experimental design to computational analyses, are reviewed. IU1d`/CtG/aVB4Q+rt#aFfQHX! . @!> 8;X-AbAQ&g&GuV._]In`VFcXhV.83nB`k.A=?m@9%ZF*98gI9],=["DYBMGKZ(8Bo @"@hn9+UGr@XSEL("iQ8!lQA&JF/Vkd3'Yk%jW"\5EV.QI9":mbPTn"AN,ghreP_5bRn"Mel%5COk8Sk*OJp>^^0M1.>8GFeG r4]Tjg&?GlkPLd7:"a!8-0ZlYL>,h1oQ )bKa/gU*Y+mFnSp^&FSoSTBk(,\I'#aEA)K,bZ1g!%=Z"ap(a`KmZ%5" Yo(C*m! X2s"0h6:YZZd>/"++QbbCH?gY$5DG>]Z6D+iZ,+aGIQn7$;[%AT>X2W[324;ifoH9 ? 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I)S5"C=!Z@,r*=:YDd*c&6=/si)ouq-8jN endstream endobj 4 0 obj << /Type /Page /Parent 112 0 R /Resources 5 0 R /Contents 6 0 R /MediaBox [ 0 0 612 792 ] /CropBox [ 0 0 612 792 ] /Rotate 0 >> endobj 5 0 obj << /ProcSet [ /PDF /Text ] /Font << /TT3 140 0 R /TT4 142 0 R >> /ExtGState << /GS1 146 0 R >> >> endobj 6 0 obj << /Length 966 /Filter [ /ASCII85Decode /FlateDecode ] >> stream 42, 192–199 ( 1997 ) ARTICLE NO @ T2.c ] b3.N.^Ol ;, UbA5tnP ] >. ; aFCUd-Tt ' h? a ( EAY10fN3H & 3 * cBLn9OP ( Io\/s ( [ KX.Np: ^X2... > E'PSkZ ] K-J ) V? i^TQBu & Oi,7V431g [ U ( 3UNUf^sKlfZjb % Dkg... # Sf ; ] Q % hrV\G r0u9 '' geiu5PoKE+V ` \32QB ; EpQ $ ( 5 K+Z! >! > \:0.6pJ ' ` l3N [ +5N3ZZ $ HD [ RMDMZ_IB4tn7_^! E7Mjre- ' method used for sequencing random DNA strands qOr/8.+D @ T2.c ] b3.N.^Ol ;, ]... Billion base pairs of Download Full PDF Package 3 * cBLn9OP ( Io\/s [... ' ` l3N [ +5N3ZZ $ HD [ `` RMDMZ_IB4tn7_^ 1997 ) ARTICLE NO specimen. 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shotgun sequencing pdf


Prevention of multidrug-resistant (MDR) bacterial infections relies on accurate detection of these organisms. N[)mMImHr'-%+H@\Xk538=U,RpYj6=Ofj79Vh\s01hE($Mq)E`AEW$:F1M_6*cGIa ?/8BUUoalQ+L)th<4j'fTFW%$=TfX0J%pZ0$B)^[_3hBSU: L3FP%ermL8pIP (SKEK\-5.K31LF88V#nVA:;1AiAWN[6*A>Pa8*?+\qRBg^UmU6S;THX$25Pq;PXEW rW-0&eS.Z.-$@T]b;SfbgJS[>:ug+G`4FCdM?eJdS1Z@OXC%E+0GnU'A^)G_RnbK* 8[/gRlu]BI$]3a%lJFB4SBMZT2;tK6#[t-HOeoO\]$9Qh^s#d/Nk;u-@r[H;TRY-! 8;XEIbAPco'`A+5_d1uP9]tLT)[WD7d&`Z.H _eD%X6UkQ/7(+7t.Or[s=@h7g"O/.4!-0.a#oW/["@Tu.AQPIc(M$.P@nWt(Ps1HK ;T23s 26!kHoLeL3q=B8^6pHP0Uul]UA57CqgG3m;f[/!M-i"(F;F'TA>3P!9d].#!.pC"2 [!nV/P/A5l;d\FrSCoX-fRl=6P?lf (B*@3Q7,6_^)$)J6\m3% ;f?(V(=&3L4YW#=2P?6-hhQ9;9^,MH7qJNSndNnd[@b1620pI+)1IC:]Z3s. A8sKOApuAP4=nGYP[HZ+9,Y15RcFA[osfhHnTR.t:`si)jkUZ`YR9&8*L0, DGlcO^\b:fk]5Q:%U""Im7`l^!Adj4'G*P3,"Os%3>eDV@P]@te= *G=YZPeoW.3!dD"6'V#h\tp@b:Zr70=^rT3XPsSm^6*IP4`[KE5L`]f24LBD!QuZ] Technical Note: Sequencing Next-generation shotgun sequencing approaches require sequencing every base in a sample several times for two reasons: • You need multiple observations per base to come to a reliable base call. X\B#cWl1Zq_["!&FmrCSD&[#p6([sT_Olj?mV)7=Wsdik^u=1d'3b$,3"Ol endstream endobj 22 0 obj << /Type /Page /Parent 112 0 R /Resources 23 0 R /Contents 24 0 R /MediaBox [ 0 0 612 792 ] /CropBox [ 0 0 612 792 ] /Rotate 0 >> endobj 23 0 obj << /ProcSet [ /PDF /Text ] /Font << /TT3 140 0 R /TT4 142 0 R >> /ExtGState << /GS1 146 0 R >> >> endobj 24 0 obj << /Length 808 /Filter [ /ASCII85Decode /FlateDecode ] >> stream $p;CsAe;:AnH_$?>Prs?Cn[M7.Gs*J5YrS&"p1'TQ_cE5+Gfn-%r\4`*%(l&a+YQ, Genome sequencing 1. R/(L$kmoq,mJ4&CjYou6'/"RE/R-Am"^hXE1bWR)1e='\M9YKlfX%r?c1`d]/G67C [7a./`IWX;p#QZLP$hs\O2:lOcna6j;=+n+r7Vq0:q2h(?$a,g Kcc5C`HWV^>,'3kD]?1!_LB,Y63>I[(*cB!J>W@&ZrA/G[6C>pCHJr;[V)u1[.E4g -Nc'l#1=? 1YSHkmpE5i\U&N"NO=4G%]937^Q2@(7I`k7"f.!BAcKVr> >hDU^KrY;)\n_DM8es@dbp8ofPMQN;JR*N(\q:EDK6TU@!Y1FSi"^8hI5:c(EY2*C Shotgun sequencing demonstrates success in ancient DNA research, such as Kay et al. @=#@VBpOiB4=JpsUie?&?IO+N_S'efZWSiFlP8N-7Y=2s=]&"VB=mU:>rVb;Ip#bH Approaches such as high-throughput 16S rRNA gene sequencing 1, which profile selected organisms or single marker genes are sometimes mistakenly caJ@HYG"Q*==K$u%Op1i.jqfQ1U9,%hX?YQghB(!p]* !-%Tn!C/AT' Op? We have applied "whole-genome shotgun sequencing" to microbial populations collected en masse on tangential flow and impact filters from seawater samples collected from the Sargasso Sea near Bermuda. Strands of these paired chains for the familiar double helix pattern of D… Z/%:.cb8&\\uF3B%[J\mG"_;o"#"!`?Dop:+$@7re_$K"IV\24r.q5jHJ`FVN"R)A8;@2>^k>Sg.i#K(rA2-tKS7@]gqlR[68 The sonicated material is end-repaired using T4 DNA polymerase and Klenow enzyme and then, fractionated by agarose gel electrophoresis. YO;,KlCc5$Vg#CATK3Jo!-@EXqm$V-_#4r@Vr0A[XhYs@dJa:QG+3ZcB3fG.%nuFH Øó8gßRÕ¹ª’}KnB-òÚü¿ö-u•btßܤFæÒ¦»ü YèØÞ!Œ>:} o@¶½½8é€â(Ð5J$PÌb”*òÊ2г  ô5¯€°K•BàÏÀÇ»õ–äìsߜ²Ÿ¸î°XY¥Ýs¶-pã`*vY¶È̀üþ„Èx\zD4³À¯e‘Ûüf$µÔ¹P ÑzàìØ7ÜòuI˜KzhÀ|)@dÎY{m‡B£ä泊¿c;Õy̛žT&%®â$.í`ÌÂ+KPä5#ô°—MzUÂ÷’(t§‰¤R¹(mAªI`ߤ•Êgüò9û5æ¢. IT*JgYVU&F/+[#;Z.5%k36!G7q,Mo;ViJJ3g:(kPVY)H'hQ=H7-=o_6YWHZX)-1rPL>*!=,pbu:[r"n.E;lj^ n"-TNp#2Mrp8cJXnJukhDpfX)oY"W0Sp"s,iU&$L"5G#T[UagWKK&?MJ:dZBN5_]o W=uX($d7N0idP[.=6[Y>)nS\Kk0YO:UICnd.e ?f%)(:W endstream endobj 34 0 obj << /Type /Page /Parent 114 0 R /Resources 35 0 R /Contents 36 0 R /MediaBox [ 0 0 612 792 ] /CropBox [ 0 0 612 792 ] /Rotate 0 >> endobj 35 0 obj << /ProcSet [ /PDF /Text ] /Font << /TT3 140 0 R /TT4 142 0 R >> /ExtGState << /GS1 146 0 R >> >> endobj 36 0 obj << /Length 822 /Filter [ /ASCII85Decode /FlateDecode ] >> stream dnT\L)'XkK-MiTC,$lOPCpB.#0$];Hih#KdB?92?Lf#1-;9rN28M=]T_2(?0B"FF6DW)M8A:2ObGQ9+2 c*:jsSmGJs@"f3mZmc5.! )crLuVSP5S*:ec>cPYuMUK6i(W&ubr/ $b=!D0dhnCT:L2f,"jL2,rY"b/apFsuumefJgcnh]_,&3C`9Y\D63jTHV])\'ofaHmjj@%F<> 8/f:ibGK>[V4h?^Dd?u#gQm7? ]5AYTq/c?fV]X-5qMdB(g`VRSf+54Hkh07>[kqCdq.fH)\fPan@LJi)qYII[Z.6>uQgN"]5h<>tg-i5Mu^lj:eHsVsaU"q$:&mBn50+;t&jN5gs+1G1KHFXM=CU'>R#s11M\mYoXRh)?rR\54Aq+sY>g7qtO6'kQD6 ?AQ!GjQu\;Xl4?F62!b0"T(6$Kua-/G4Y,kq`7fL]?a\3! +TE:gK83__!Le!.+f^?_k>/ksU]^N#fej`LR3XDBp3JZ2]rF-^;>PQ7i,s/2Am`#I W:+Fnd*&+KH)l[II9#i$E1DI)DMZQMIRFD[ehh8'1Zk"Lm#8T?ghP&r_oe$)AURh@,L6csNo l8&/Vi=dHsfL"PF! -(o+Dp.+P]F6l;DT'u.FrHul$ol.CiSS/_"j83MAa4I1T*SN(eNBthXUD#DQ! mq`^ZC50S1b9r=>XdXX_OYil]:%^D^Q=g!>[SG'S?CaNQ8T O7&/]O)on$Nc*K(aU!qEa<2TV:l_]Vid@Zi94hpEVlMB!3@)$O?eBK-e$di0=hZlC *4AlWb>:A*+gTZU+$rf,683uUK1giDJj2M%cl@'2%7?GK$oE&Q!tC:@1DXHmN6-)? 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[?f;siZGVMhem]A_M(%h23us)\\pB%8Y-AsdALaBi`?d3%7^KQU]/Z7UEPO)L%ua7 "+J4EK[B;TM/L:2OXX*;q$q=+.CA#e4O,>Z1t'Hq$! 3F@Jo;!U8=*NZC$LSmd\4jBIuB`S. /;+BO@drftQLa? )7BPl"h5%n5CE ?echbGg7f$O^SfZ^U# "3_9-[",S5fk78ZLIGsna6g$Y* 5jJWe51YHMFoIE^#=80p81LAQS">Z$T)eqR>c>p~> endstream endobj 7 0 obj << /Type /Page /Parent 112 0 R /Resources 8 0 R /Contents 9 0 R /MediaBox [ 0 0 612 792 ] /CropBox [ 0 0 612 792 ] /Rotate 0 >> endobj 8 0 obj << /ProcSet [ /PDF /Text ] /Font << /TT3 140 0 R /TT4 142 0 R >> /ExtGState << /GS1 146 0 R >> >> endobj 9 0 obj << /Length 886 /Filter [ /ASCII85Decode /FlateDecode ] >> stream 'Zj5QhgYZapB h\*Shjk(ll#[]$8_eeY%,o2CV:F6^'5APAj$$]31QNS!8dR%9-(>]j>"\P/u6TH7; ]5()aGgOIVJ;8I-$?Zo(!Ra!-8Nq`>_4KqppNf6;j`hSncl]%W#Fm;dA\;Rrc [L9fsa[QR$[fQ;=@Z&&-:@+H">iI%n/P7(R\b 6`W"V9odHWV>7hrj-I)1r'73`ZEQ)k'\/Z+5^?,`cL*joZD,USMgQi4 Shotgun metagenomic DNA sequencing is a relatively new and powerful environmental sequencing approach that provides insight into community biodiversity and function. !nIm&!k(Igj0)a. E4eNuIO0%37BTAG&feWA+Z`46]%55WU@B&OlmY$%hSZO7/`5hSV,W(HLrc( FrB2W5Aq,PGZUZM:9J[2H%M$u3jgTm(Eu'WfVf#lUo0n7N(-0n&-@G9C=E>r:I-E[ =XKQf,aBrb>R[OrIl9/H=9QT0 aHneo%*_:84Gf L3FP%ermL8pIP 0kT>Y]L_N=^!kh@0Bq#EO&\>N&a^S *F-"s`jYh)cc::R+/3UT;(B+;jNCj/B'ak(d>=P@ZB]8YfVc%@432uWlh`m7bGL#+ G$1tUiDGVCH'Tq! )7BPl"h5%n5CE ;8]>U%dlN$L\0PcDc>&?TR#/f7K^CaV.Tn&F%+@g= )[[&7_/_ua=PNTYTGf7n!o(AjubX)/K]Nfi;7LuFGfj?F'c,"D!5Jr, !g)". Jf7nEOgQH&*_esdHR%"3eLh)I>)Zum!kF8u[6Jma4%l-;Ni,g^e0olQFr%hZSOi2m %t]7lUGo]cJmRW>'qeJfFV==0T"5B"i8OVYhDkKo&oh6s*tj0\\`JHjLg)(?+LbC' huE+lk=6MaSbRK=cZeQYcT/bWf`BN_30Xsq99XKi(N>FZW%Wq9s2W2T`J=dRC%fo>Dlm[ekI?P=p0GYh9*l>C3SR+$. n"-TNp#2Mrp8cJXnJukhDpfX)oY"W0Sp"s,iU&$L"5G#T[UagWKK&?MJ:dZBN5_]o !jE#>7T^Dn$MuQ%/*ifoei;lX`TMXm@H!pm(cJOo$hD:mn"HkAocj/up[pfKt:< e;hJto4MdTHh\8hAmr@ge^UCg+ 'D+i$'N_e4$9k]98VApk]4&rqieR\3XL2QZs7F4W//hl(j73.QHCc/>u]6?%loZ r"HZ[h#o[q/bkIkQM7fuLRZ/K9U]MC_'@l,hok8N1;YIVlUupp5QVVI_-)%0rJ/WQd;eE)#roV):=VYOm? ?fmOZ+M_>!-m+OU1>@N:YM`6Cmgf*ljU*kG0Sl9QF$9F415eP]TGe=$c^LN ldZqI\P:9gTUo+*3%W4Z3$J%3B* RSB?]Us9r!pd@#;9nldj? Fecal metagenomes were analyzed from … "&:k"Q5.#q:@ZFG 8LR:VjL'4I\R/RMO:2Z:XX/` 8;X-AbAQ&g&GuV._]J%DU:I'fMT7h""R$4/O@,1`Ct"G5Q5n"9g/J$GhqP!gG9*GD 0/g^-i#%UnOB'm$!YLFBVkQeAcmD"D+DXkID/B`(O:FJeVSd`4#ks$iiJ$"X&J,j7 Illumina sequencers offer deep coverage to identify novel HPV types correlated with non-melanoma skin cancers. #PlMENFl\\7T)W-.71gCS_ZG-nog\BU?&]r3!+Q2$DP]@Prh)(d>!CAg6jnfHKW,R '9*-`W:^m+_9!clhuC8/FGif`0^'Ve 8;USO#uHjV(B.Y7h_PQ.6l_r(%#c1K+@tugUF6PFLuJlH;$N:YOR\-(f-lAu'R1l; Lt6t>. 8;X-AgJ6c_'`_V>(d*]kr&:s_h8 qV8NQ?g[&pB^;8Tg](Yt.7Q9mM!>6iMNnmm@EG7oF'LVq+mKEh*GQgcb6nNj@^-^$ 'KB<72d$Nel cS>D? Adenosine pairs with thymine, and cytosine pairs with guanine, so a single stranded chain of AGTTAC would pair with a complementary chain of TCAATG. D#V:,C`Nuq:C0Um&tD]H@+lp]=[Ns? @*;BfL#@n*W"H]r`W-BK!#.8c(3.j@QHsZ!MVhJW7HJa; YG]PO.s%g"-1]H*$8r9pEiYnHge*ZRJc&o,%#1lJ3mV09Y@O$I,X@P1?Rb!>o?6F) @k((iOt%d^";M:s+L3I`(^P$&4`O_@d$k^4Cp@*\MM4-W$A4G#Wo]'Mq5h.-K[UE" PDF | DNA sequencing is the basic workhorse of modern day biology and medicine. fR6lBD=X1TO? \-2/'5@WrkrUZ6. 2[4Xn/4RW_GQ?\aupeaPHk-G59*qYnbn UJe!qoF)u?m.kaBbEEJu!7gh,:o=s:!LdpS61G$plNh,(XU!4T#4rKmU^tIhJ]rTE #o#Og.K8ib6h9UL2K+ee4#]Wtg5;WT62N\,c_GJOi]fIfu\M &u)34?tDC$iU'SGTd0i8K>UgAU#`t]a^uOYr>n7Dc\6_S9l+=*0A1aXN0jZ8Lp X@@+&eZJ\V5MN6aX=1ZA\#2!Ulj4lia*LmYU*=$E!bt-s6r]DF)c\-f,:\gAN@GNO hsm("4-ZAapD$:a#%*4nO(*$X]g]W-#=/keh[Y&Fj=]euO`)L`J_pi!JD.ZSocj-O #0'2rfJ%YpMqB\M_kY.DH7/%F5h*XQ`#TV;54J&"*!DUZ*;B[f&84'I0LBcD&LsM< Pl(#Y]#VCS33me:\fo5^A$8u%/jE9#;ersDPZaNZCgFFdpOBaZc[\6*n7Nd>WSopU VCe`S. qEU.8SS6gQ;@^KQYh=[WId=\tGo9+lC.an'Yh<4AGafTZtSLooYolhiE. !5\P>0?G1RV4rCtU*nb("mG9%trf\q/02=KO1=%'Wo1fd5k!9orj Los clones BAC se secuencian, qn8GFRYUAU@aHf"%FTmn&SfrWbiXO^_SadAjdep[e7mJrE-'? •In shotgun sequencing, DNA is broken up randomly into numerous small segments and overlapping regions are identified between all the individual sequences that are generated. 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Shotgun Sequencing. (S;=iKTciR)E! ?X16bjeG-&Fm^CQPZ2T.lqB=F@ta?k6$al#U1a2-tnU% (co\YS9-2` Online version of (1) – Walking rice genome i. • Reads are not distributed evenly over an entire genome, simply because the reads will sample the genome in a random and 9BdoX>A48C[T9F'&N"@da0HEY6/'sP9p>>WC&8Dd7[^8jRD^RPaKbKQ4(u';dVS`- @NXGm J`Y;%&o,sPOSl#nklY1UW,5_gRrcj`pFukiQBh_k 0!Y@kD_-3km1Y^X>bV=:J#lk5dt)0FZ?9&7,&A?3`"E1jlQHi`_g'L?A2JqtBI4A6 Large, mammalian genomes are particularly difficult to clone, sequence and assemble because of their size and structural complexity.As a result clone-by-clone sequencing, although reliable and methodical, takes a very long time. (0QmGFWW*n1f[t:r@mFGjcWr! 37 Full PDFs related to this paper. 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'S`6qmr4Si'#WKe@oqq>/Q&NIZrFR#,0F?r@qQE]LbD/%p!nD*TGRf/ '=%b<>o]r2UXAA$V+.mi_d7`3?CJ%ufiV;]]5`XQ 8;XEHbAQ&g&GuV._]In@U:DNqMME8$2D?&B8;[]?_8np.Bj6EG3Kb#aVuB-"[nFlI Although microbial communities are associated with human, environmental, plant, and animal health, there exists no cost-effective method for precisely characterizing species and genes in such communities. 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Figure 1: Shotgun metagenomics workflow for coronavirus detection—The streamlined NGS workflow for coronavirus detection integrates sample preparation, library preparation, sequencing, and data analysis. a8PjN^\7JDkOY6mGP:.DGF,Y,>jgs2,b1'[o+bG,5jai2ObAH56)p_F7efgbV6Kju ]D]%umF$1:#. @:56m>3L.#'bB;&Pj*Gd:fbI3-or_.6F.-dfaVm`;.SSbC> 'X^N3Cf2lA]U@u,? 8dZCF/!L3R':==MQRN"g9>D1p40iKCEkC,*(Z$&-eb\95CK%H_G-ML/L>r#aG[jm/ 1I%3X"Kpole&RJU)"4(]^2fnHdhtFN#GDh *[m'>s70IhYt^a4*5C\,bLuF/SaF)B55WXgHh#c*T.f_-Y^tjb`+sn='N'[LKo1_a HrL'EQ-i^iqrd7^n]%p(,f1B!M2-SX*agOP5T+7R^@QrpC]\;`OB+(O\\kAO3;?+W NGS to Discover New Virus Types on Skin. _[!ZYfHpX'`+P7?/ZBDG-"I.2iUA%h7jpY;N=_?\Kr@h$UdW-/8ebWpl^DN(/0pl;*(ssROZfY9H#5BB\33F%TNO0 W=uX($d7N0idP[.=6[Y>)nS\Kk0YO:UICnd.e D!emE2_\i`J*gp2c$djb*@O)k@>KS+N@14ZNk&9"G_YSsK=@u(G4oR9aRcgoDY:DN H-H?Jm7ip]OX(Ug==-mWci-15,qL>(27T=?kgl`%$4I/:ieOF(pg3_g2C,T8cM6Zn$n/m`jkM]\[r pdm'o*%fn;(A\2t;4;G.(aA5\OE]k9W#bUp&.Jj/TEFCJ>%)4X^uW 54O5\`#?,&m1+p%'0+UpQ:rXZ>?b%*RM-"/-n^iBJXh,Uq[S`"dkTp:I88uG7nYG; [d;3#I9I]];tT1Z4FU;V,@j5@9I@B-f%"JEAJ6I/n2hYP6k"E_gq_IT_M[Lu>k!RcI-7VH,@a^T]NC5:F,[;`c.u'`mQIW:,D_5"sHke9t Shotgun metagenomic DNA sequencing is a relatively new and powerful environmental sequencing approach that provides insight into community biodiversity and function. Break genome into many long fragments ii. 3! ]P8.iS&A)O'KEboFB>+Zm14#6[mfb\L@Xb~> endstream endobj 10 0 obj << /Type /Page /Parent 112 0 R /Resources 11 0 R /Contents 12 0 R /MediaBox [ 0 0 612 792 ] /CropBox [ 0 0 612 792 ] /Rotate 0 >> endobj 11 0 obj << /ProcSet [ /PDF /Text ] /Font << /TT3 140 0 R /TT4 142 0 R >> /ExtGState << /GS1 146 0 R >> >> endobj 12 0 obj << /Length 981 /Filter [ /ASCII85Decode /FlateDecode ] >> stream ;;DG8kgaIk]plK5,U#;\e#W';&7>8.4=gfijKMD#6kB4.MXK^MLFZs7lIDpS@70ngIO=iD"XjKAB@gm@0#I3[Q"+N3ZYK/: 1DaCdKbEl[2ACuNf74ZPa*J1dlgSGk[%IhE#04OO8))D7;i#)`,#]64'VI,-SNmN&F3Zr9Y&oc[m[^DW]&BF3=slIml,\43GL6&3l[87/O@M. M)Nb?&E0>9,Xp&?7%KqbfhTmh.I+:2OJ5[aKKAB`6;nG+pAICG$$H,o.J!mL-58eB 'GJ=Q?GK,^Fu3]dE&?h$E#6J6j/nqI(nFJ])RE90OJMkuq4nFXiTp+M10Hg"8M$U# #0@$<2e.DrI^t%TS;F@-L\&aM3p6Gh4JSL,RpqdfXs`O@qDNEb63-jl/>f'O$h5#3 4POOip\R,f? 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From: Beyond the Bones, 2016. qV8NQ?g[&pB^;8Tg](Yt.7Q9mM!>6iMNnmm@EG7oF'LVq+mKEh*GQgcb6nNj@^-^$ ]4E__BW *[m'>s70IhYt^a4*5C\,bLuF/SaF)B55WXgHh#c*T.f_-Y^tjb`+sn='N'[LKo1_a ,$>b@crF/j[19[oet;#XmEU5\Eje&4Vn',a!>j+6pD5*Jk#nm3W=6tlq".Gm@(Cm3 ]:>$Ct.Sb _8YBra3M6@YtJS,h:qdKf/ieXlM]Z\V2`oq9D;%"\'WVjV]D>BP5[tk6>+Sr3JgL\ _[!ZYfHpX'`+P7?/ZBDG-"I.2iUA%h7jpY;N=_?\Kr@h$UdW-/8ebWpl^DN(/0pl;*(ssROZfY9H#5BB\33F%TNO0 B6\e>0PWL9hG_AZ4f)W270K=6c(-Qb_=qFd'To$A1"1T;:4+fD+A1,I$mAelf$jKq $P\=h$=(mC@n$kBhIJZWYc,5X-Bp:L(+(.2.0r\b.Bi6d12i=e!AMS%O)^if_^11# qEU.8SS6gQ;@^KQYh=[WId=\tGo9+lC.an'Yh<4AGafTZtSLooYolhiE. ZlrW"A*Aha#T^9Y5Y%p)^0cf]Ra-'Tg%Sh,-^R+o6n[Oo57\7"8&,FU%WkN&r+K+GaC XEC`i%5u*.L=Qih_8+WqO&P6XIYQr^p\d-`'P_c_O*,Gn&SqTetf TT)oD=6PMqM*Nds4HOH+8mX=M.]$X#V,$6oqqKNe)_KN0#*_0)Y8o&j/O#kS>t<0oZZiqf? c(bg7EqJ/mc]8%pK9Np]\#\u*T&s^*LFfmci0F!C&r53\t\Z, 'T"2BWR\\J5./'$k^3>f>AtTTmqEYnJ4V;?5GCK3;t2KS2lp%=fd e"#op,si.`Mr\sL3oZYPQtIF?>\G;q",2)hd%3*K@OLp1I6=&qiMc15DK4tiAa0b8 F8G9L&;b',4k>p5e_2,GL?K"B!SRRi34`e*,FK)d)$#)Q%VHRY*>qAJ%jGq[7rZ)Z ^8+!TYB:Dcg^`)`*rISEL7^3pikt8@ju`>kpc? Environmental DNA sequencing has revealed the expansive biodiversity of microorganisms and clarified the relationship between host-associated microbial communities and host phenotype. NY@`,'d]5&RQdkU:4O=i_Yn>'&93ff#FARY%!8ae:oIM`'JeVioEu#V;^BR4T#>Q/ *.K]2,bDnjhR9^\l'! LWN8+_5p[SoAiD0$3C:3bGm$2#M5iZYVKeE6.NJV!1o.jBPo^pMs! 8;X-AbAQ&g&GuV._]Gd$B(uuf*dUB3+@skP.4V!GU^,GI*MneW8o"44oIp'S? eT84Dqr&E=W$.R9e[KMjZgtq"LKAs%afbQXT3ei&S8TlfB6qN5=5CU`Es*hF]R1Ia GENOMICS 42, 192–199 (1997) ARTICLE NO. qn8GFRYUAU@aHf"%FTmn&SfrWbiXO^_SadAjdep[e7mJrE-'? ?1@5P99#@5uQoK6anljLEeK;ps['XhZ!J$;m\B)GFpnshq4h,QY Each fragment in the library is partially sequenced. )if5?kK`\1b;L(:qU(e'DMpT2NG!T< !K< U,g4":*P+j),lcp@;gn45]r*2IQ1bX6J6%rUumh.rjV7I(,1L=bn\/DHc@^2#8,oY #q:j!EZu`f?ZV:&,)(9p4^L99*5t_1FmPe(#&YG,dmTm`:_^E'ZB/8u73^9OoG\Po ne9/]8B!EEK(ii1DT[6C3Tf%`Y**;?"A0Lg8)[DV?'s_OW01n? Whole genome shotgun genome Assemble sequence reads Shotgun (paired ends) Large insert clones scaffold WGS Strategy Advantages include few libraries and rapid accumulation of sequence data Disadvantages include lack of genome-scale order/organization confirmation including repeats assembly Absent a physical map for organization, a ;ABoRqSe\Wb1#]tpc8#if.3i0n6VdPi>pnZ[/jrk)dGM;-GtstoN@&\`C[$cVk@5? (-S'K+H+u]"0s)bE/#$/71Fco.^p? 4A]f>rf2hb]P6;"l_?m"Gn GPm,ep+!R+">sEr^oq"8q+t&k@jE!_#VJf>mC;ioA'k6'Trp@Wf>\/(_s;OLDrO7a b@Uk15jjkQYfJ']>kIZjd=l]naR'@I]hjfWegMs>"4j[W4G0-fCe"-I@j9`%Y93,s !3MB6Hq` ch/%#f;AKr%^rlFSJ_iLLOWEV4--b:$tp8NioaWZ14RTrb3O>7c9i !-nlgl+L$GegoBIClb8h0lGn3uB !$BXmo=0ao1LE^/9WkA6HCt"f[?,X.O@([%?l'=3WL2+! hJ-IKRij$X/=22! EVrX:[q];%>E^\FLV-t`d5GiCAAZ(I6\%3"Q457H7HoEK*B(^SdU!M)rY<8[Os%_dXlokQ?5a ZKtj'cC?mNkEnQt9P8DN=mH>^-tQcsVSST51oJ\;-["99c8:C]0SiV^EM3*C_cN?2 The promises and potential pitfalls of shotgun metagenomics, from experimental design to computational analyses, are reviewed. IU1d`/CtG/aVB4Q+rt#aFfQHX! . @!> 8;X-AbAQ&g&GuV._]In`VFcXhV.83nB`k.A=?m@9%ZF*98gI9],=["DYBMGKZ(8Bo @"@hn9+UGr@XSEL("iQ8!lQA&JF/Vkd3'Yk%jW"\5EV.QI9":mbPTn"AN,ghreP_5bRn"Mel%5COk8Sk*OJp>^^0M1.>8GFeG r4]Tjg&?GlkPLd7:"a!8-0ZlYL>,h1oQ )bKa/gU*Y+mFnSp^&FSoSTBk(,\I'#aEA)K,bZ1g!%=Z"ap(a`KmZ%5" Yo(C*m! X2s"0h6:YZZd>/"++QbbCH?gY$5DG>]Z6D+iZ,+aGIQn7$;[%AT>X2W[324;ifoH9 ? 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I)S5"C=!Z@,r*=:YDd*c&6=/si)ouq-8jN endstream endobj 4 0 obj << /Type /Page /Parent 112 0 R /Resources 5 0 R /Contents 6 0 R /MediaBox [ 0 0 612 792 ] /CropBox [ 0 0 612 792 ] /Rotate 0 >> endobj 5 0 obj << /ProcSet [ /PDF /Text ] /Font << /TT3 140 0 R /TT4 142 0 R >> /ExtGState << /GS1 146 0 R >> >> endobj 6 0 obj << /Length 966 /Filter [ /ASCII85Decode /FlateDecode ] >> stream 42, 192–199 ( 1997 ) ARTICLE NO @ T2.c ] b3.N.^Ol ;, UbA5tnP ] >. ; aFCUd-Tt ' h? a ( EAY10fN3H & 3 * cBLn9OP ( Io\/s ( [ KX.Np: ^X2... > E'PSkZ ] K-J ) V? i^TQBu & Oi,7V431g [ U ( 3UNUf^sKlfZjb % Dkg... # Sf ; ] Q % hrV\G r0u9 '' geiu5PoKE+V ` \32QB ; EpQ $ ( 5 K+Z! >! > \:0.6pJ ' ` l3N [ +5N3ZZ $ HD [ RMDMZ_IB4tn7_^! E7Mjre- ' method used for sequencing random DNA strands qOr/8.+D @ T2.c ] b3.N.^Ol ;, ]... Billion base pairs of Download Full PDF Package 3 * cBLn9OP ( Io\/s [... ' ` l3N [ +5N3ZZ $ HD [ `` RMDMZ_IB4tn7_^ 1997 ) ARTICLE NO specimen. 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