1/20
`
`10%
`
`"N
`
`{is DNA/RNA
`
`SS DNA/RNA
`
`“Ci/i Denaturatmn
`
`149/”:
`
`390mm Adapter Ligatian
`
`
`
`1594 Amphfacatmn
`
`
`FEG.
`
`”E
`
`
`
`10%
`
`"N
`
`{is DNA/RNA
`
`SS DNA/RNA
`
`“Ci/i Denaturatmn
`
`149/”:
`
`390mm Adapter Ligatian
`
`
`
`1594 Amphfacatmn
`
`
`FEG.
`
`”E
`
`
`
`2/20
`
`(335 DNA/RNA
`
`SS DNA/RNA
`
`i Denaturatian
`
`SS DNAI'RNA
`
`i
`
`First Adapter Primer Anneafling
`
`..
`
`....
`
`Primer Extension
`
` i
`
`N1,N1!6?C-Wm
`
`,5, Anneaiing + Primer Extengion
`
`N1,N1,N‘E
`
`
`
`Dispiaced
`Ampiificatian'
`~
`
`’
`'
`
` .
`
`HG. 2
`
`
`
`3/20
`
`(335 DNA/RNA
`
`SS DNNRNA
`
`i Denaturatian
`
`SS DNAI'RNA
`
`Primer Extensien
`
`with a Nonutempflaied Paiymerase
`
`i Second Adapier Primer Extensim
`
`
`
`i Ampiificaiian
`
`
`HQ. 3
`
`
`
`4/20
`
`(335 DNA/RNA
`
`SS DNNRNA
`
`i Denaturation
`
`SS DNAI'RNA
`
`iLigate Second Adapter
`
`
`
`i Ampiificaiian
`
`
`HQ. 4
`
`
`
`5/20
`
`(335 DNA/RNA
`
`SS DNA/RNA
`
`i Denaturatian
`
`SS DNAI'RNA
`
`i
`
`Primer Extension with SEX/EARTH RT 0r Bst 2,12)
`
`N1, N1! EEC-W"m
`
`,5, Second Adapter Primer Anneaiing + Extensian
`
`NN1,N1,etC.W
`N2, N2, etc.
`
`
`~. _ Dispiaced
`Ampiificatia
`
`"
`
`
`N1, N1
`
`HG. 5
`
`
`
`6/20
`
`i Step 4. Semnd Adapter Ligation
`
`CES DNA
`
`
`
`i Step 5“ Ampiificaticn
`
`
`
`PEG. 6
`
`
`
`37/20
`
`gggggx..............000iiiii
`
`wwmu
`
`
`
`
`
`......................................0,meaaaaa
`
`
`
`@QQ,,,,,
`D00aaaaa
`
`
`
`Emmfiafia.wA.
`
`
`
`
`
`www“unnucammmflKama/qvceumm.m
`
`
`
`
`
`
`
`
`
`mmmfififiaqEmmcmgmnéxza£2,cammcfixmESE.m
`
`
`
` Ammmhmfiaem{Em9wEmrmdv.<
`
`camfifimcmmaw
`
`<zmmm<zmmu
`
`<zmmm42mmm
`
` gamma;
`
`hmwgmmxfiga.mw
`
`w@fi
`
`
`..................................................DUO
`
`
`
`D00
`
`
`
`
`
`
`
`mmmwmtuwcmbmwh®>mfifit};EOmemwxm(“manta4m.
`
`
`
`
`
`cm5E<2m:3me
`
`
`
`
`
`
`
`
`
`8/20
`
`
`
`.......................................@mm88888
`
`
`
`EEQEEE.m,A,
`
`m@E
`
`
`
`mmEmEfiomEmwcmgmma‘azm3:3:Qmmcmwxm5E5.m
`
`
`
`mmmwmtmmcgmfimmfihmam
`
`.w.wwwaaaaafl
`55555,m.cowmmfl5%”quwgmumw
`
`
`
`
`
`
`
`.......................................000
`
`
`
`000
`
`000
`
`Ccmwwhswwfimfl9%
`
`<20mm
`
`<23mm
`
`42%mm
`
`42%mm
`
` commmfl
`
`hmfimfixWEE.Nfl,
`
`
`
`
`
`
`
`
`9/20
`
`(is RNA
`
`
`(is RNA
`
`SS RNA
`
`i 1.
`
`ds RNA
`
`SS RNA
`
`mm i 2.
`
`.............
`
`{is RNA
`
`SS RNA
`
`
`
`10/20
`
`BE
`
`CZ
`
`532
`
`E2
`
`F2
`
`$2
`
`
`
`was(DC)CECE
`
`N CI? C)
`
`
`
`PEG. 1%
`
`
`
`mww.anm43%.anm
`
`
`
`
`
`FEG. "i2
`
`
`
`13/20
`
`G1
`
`Nu;CBC)CDC) C)
`
`U? C) CD
`
`A C) C)
`
`L9 0 (:3
`
`N C) CD
`
`
`
`
`
`
`
`
`
`
`
`14/20
`
`mayWaP!mEhneGuNinw
`fl“
`”m”rI..\W,mmLAANDSS
`
`an
`
`M“,a.D
`\n/WI.3bilk
`
`500%
`
`
`
`
`
`€mmmma-m:QmmE3%Exammagmaumwmmmfihcz
`
`459G~mmmmmmmmmmmmmwnm
`
`4600
`
`350%
`
`300G
`
`500‘
`
`GOG
`
`50G
`
`000
`
`
`
`g1”,1/17.1/17.1/174.,eré.aw.
`
`--fl0%30
`
`HG. ”E4
`
`
`
`
`
`
`
`15/20
`
`
`i-Nuaen Library (39ft bar)
`mwsDNA Library (righi bar)
`
`
`
`
`
`(3-) C3
`
`N U"!
`
`N C)
`
`
`
`
`
`
`
`
`m; U"!
`
`m; C)
`
`
`
`
`
`NarmaiizedUniqueReadGaunt{permiiiianqupasssd}
`
`
`
`
`
`
`
`16/20
`
`C) Human chr21
`
`@ P0010000
`100000 ————————————————————————————————————————————————
`
` G
`
`100
`
`150
`
`
`
`Read(3000?:
`
`56
`
`Fragment Length Bin {Esp}
`
`E:E{3, 1€3
`
`
`
`17/20
`
`nascent
`
`(gmwing)
`strand
`
`paiymeraae
`
`5'-
`
`“5'
`
`NNNNN
`
`tempflaie
`strand
`
`HQ. ”$17
`
`
`
`18/20
`
`“30 :5;
`
`
`
`ReiativeCount
`
`W 2::
`
`1042 333
`
`1QQ4ZIZ"“‘
`
`1045
`
`”E
`
`.
`2
`
`.
`3
`
`4
`
`:
`5
`
`.
`6
`
`:
`7
`
`8
`
`Number Of Nanntempiated Eases at 329% M the First Strand (at)
`
`HQ. 38
`
`
`
`19/20
`
`.._~~1925
`
`\_ ~1929
`
`
`
`20/20
`
`DNA/RNA
`
`5i
`
`3!
`
`13’ - Ligaticn
`
`DNA
`
`..
`RNA
`.WW
`
`
`
`i
`
`3.
`
`
`
`
`
`mm NNNN
`
`RNA
`
`
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