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  • AAAAAA.. TTTTTTT.. heterodimers

    Good evening all,

    I have prepared (ds)cDNA from viral RNA extracted from plasma that was subsequently sequenced on a MiSEQ following standard library preparation.

    When I got the reads back, I only have TTTTTTTTTTTTTT or AAAAAAAAA sequences for 99.9% of my reads. Read have good phred score and are of the expected length (100 bp).

    Any ideas about why I might get such a result?

    Thanks!

    Nicolas
    Nicolas Tremblay
    Graduate Student

    Cardiovascular Genetics - Andelfinger Lab
    CHU Ste-Justine Research Center

  • #2
    Could you post full sequence of few reads (both reads if sequenced paired end).

    Comment


    • #3
      Hi, here is the sequence from a few reads:

      @M02846:68:000000000-C39D4:1:1101:14422:2079 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>AFGGGGEGGGGGGGGGGGGGGGGGGGGGGGFGGGGGGGGGGGGGGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFABFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF
      @M02846:68:000000000-C39D4:1:1101:14422:2079 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected][email protected]@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@@@[email protected][email protected]<@@@@@@@@[email protected][email protected]@[email protected]@@@@@@@@@@[email protected]@@@@[email protected]@;;;=?;[email protected]@@[email protected]
      @M02846:68:000000000-C39D4:1:1101:14218:2110 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>[email protected]@@/<[email protected]@CDGCBF?9BDAB;DFFFFFFBFFFFBA=AFFFFAFFBBFBFFFAAFFFAF;FFFDFFFFFFBD=DDF-9>[email protected][email protected]@BDF;[email protected];9=
      @M02846:68:000000000-C39D4:1:1101:14218:2110 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@;[email protected]>[email protected]@;@
      @M02846:68:000000000-C39D4:1:1101:13886:2230 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>AGGGGGGGGEEFGGGGFE/@[email protected][email protected]>DFFFAA;@[email protected]=FBFAABCF;BDDDC-B;-B;FD;>[email protected]@[email protected][email protected]
      @M02846:68:000000000-C39D4:1:1101:13886:2230 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@@[email protected]@@[email protected]@@@
      @M02846:68:000000000-C39D4:1:1101:12194:2260 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>AFGGGGGGGGGGGCGGGGGGGGGGGGGGGGGGGFFGFGGGGGGGGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFAAFFFFFA=DFFFA;FF;AFFFFFFFFFFFAFF;;ADAFFAFCFFFFAC;C
      @M02846:68:000000000-C39D4:1:1101:12194:2260 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@
      @M02846:68:000000000-C39D4:1:1101:12722:2310 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>[email protected]/[email protected]@@[email protected]?BBBBB;@B?;[email protected][email protected]?B?B?B;>[email protected]>;-;BBB99B-B;[email protected];[email protected];=>@;[email protected]
      @M02846:68:000000000-C39D4:1:1101:12722:2310 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@[email protected][email protected]@@@@@@@@[email protected]@@@[email protected]@@@@@@@@@@@@@@@@@[email protected]@@@[email protected]@@@@@@[email protected]@@
      @M02846:68:000000000-C39D4:1:1101:20711:2313 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>AGGGGGGGGGGGGGGGGGGGCFGFGGGGCCGGGGGGGGGGGGGGGFAFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFBFFFFFFFFCAFFFFFFCFFFFFFFFFFFFFFFFFFAFFFFFAFFFFFF
      @M02846:68:000000000-C39D4:1:1101:20711:2313 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@
      @M02846:68:000000000-C39D4:1:1101:17123:2486 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTATTGTTTTGGTGGGGTTTTTTGTGACTTCGACACTGTTT
      +
      >BBGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFDFA=D----.0000://.-<..--.-...--</00000....-:0000
      @M02846:68:000000000-C39D4:1:1101:17123:2486 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@<[email protected]@@[email protected]@[email protected]@[email protected][email protected]@[email protected];[email protected]@;=->[email protected]?;[email protected]<=;[email protected]@-
      @M02846:68:000000000-C39D4:1:1101:19948:2714 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTATAATTTTAATATTTTCGGGTGGGCTATACAACTGCCTTTTTCCTTCTTTTCCT
      +
      >>[email protected]@@GG<FFFFFFFFFFFFFFFFFFFFFFFDFFFFFAFFFFAF---9.////9///////;///--..---.;.///////;/////;/9/://;/////;
      @M02846:68:000000000-C39D4:1:1101:19948:2714 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAACAAAAAAACAAAAAAAAAAAAAAAAAAAAAAATAAAAAAAAACAAACC
      +
      [email protected]@@@@@@@@@@@[email protected]@@@@@@@@@[email protected][email protected]@@@[email protected][email protected]@[email protected]?<@@--9A--;@-9-;A-=---9-99;@---9-9-;-;////----------
      @M02846:68:000000000-C39D4:1:1101:14946:3025 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >AAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF
      @M02846:68:000000000-C39D4:1:1101:14946:3025 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
      @M02846:68:000000000-C39D4:1:1101:15839:3063 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      3>>[email protected]@BBBB??B<BBBBBBB?;[email protected]?B<[email protected];BB?BB;B??BB?>=?;[email protected]@??BBBBB?--BB?-BBB?
      @M02846:68:000000000-C39D4:1:1101:15839:3063 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@[email protected]@@@@@@@
      @M02846:68:000000000-C39D4:1:1101:10519:3107 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>AGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF;BFFFFFFFFAFAFFFF
      @M02846:68:000000000-C39D4:1:1101:10519:3107 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@[email protected]@@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@[email protected]@@@@@@@@@@@@@@@@[email protected]@@@@@@[email protected][email protected]@@@@[email protected]@@@
      @M02846:68:000000000-C39D4:1:1101:14081:3115 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>AGGGGGGGEGGGGGCGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGFAFFFFADFFFFFFFFFFFFFFFFFAFFFAFFFFFFFFFABFAFAFFFFFAF;DFFFFFFFAFAFFFAFFFAFFFFFFFFFAFFAFFAACFFACC
      @M02846:68:000000000-C39D4:1:1101:14081:3115 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
      @M02846:68:000000000-C39D4:1:1101:10255:3290 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>[email protected]FFFFFFFFFFFFFFFFAFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFAFAFFFACFFFFFFFFFAFFF
      @M02846:68:000000000-C39D4:1:1101:10255:3290 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
      @M02846:68:000000000-C39D4:1:1101:20826:3326 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >AAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF
      @M02846:68:000000000-C39D4:1:1101:20826:3326 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
      @M02846:68:000000000-C39D4:1:1101:9032:3354 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>AGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFAFFFFFFFFAFFFFFFFAFFFFFFFFFFFFFFFFCAFF
      @M02846:68:000000000-C39D4:1:1101:9032:3354 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
      @M02846:68:000000000-C39D4:1:1101:21178:3402 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>AFGFGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGFFFFFFFFFFFFFFFFFFFFFFFAFFFFFFFFFFFFFFFFFFFFAAFFFFFFFFFFF;DACFFFFFFFFFFF-DFFFFAAFFAFFFFFFFFFBFF
      @M02846:68:000000000-C39D4:1:1101:21178:3402 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@
      @M02846:68:000000000-C39D4:1:1101:18043:3408 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      3>AEGGGGGGGGGGGGGGGGGGGCGGGGGGGGGGGGGGGGGGGGGGGFFFFFFFFFFF-BBFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFAFCFAA;BFFFFFFFFFFFFFFFFFFFFFFAAB;@B--;--;;-;
      @M02846:68:000000000-C39D4:1:1101:18043:3408 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@[email protected]@@@@@[email protected][email protected]@@@@@@@@@
      @M02846:68:000000000-C39D4:1:1101:12925:3455 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTGTATACTTAATTTCTTTGAGTGTCTGAGTAATGTCTGACACAGCCACCTTATTATT
      +
      >>[email protected]@[email protected]@GBBBBBB?BBBBB??<[email protected]?BBBBB?BB?B;B=BB;B->-9/////////////////.;///9//;///////////9/...9....///;9/9
      @M02846:68:000000000-C39D4:1:1101:12925:3455 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGTA
      +
      [email protected]@@@@@@@@@@[email protected]@@@<@@@[email protected]@@@@@[email protected]@@@@@<@[email protected]@?;[email protected][email protected]@[email protected];@@[email protected]@----------9---9-;--;------------9--//
      @M02846:68:000000000-C39D4:1:1101:23101:3568 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>[email protected]=BFFF-DFBFAFFFFFFFFBFADFFFFFFFDAFFFFFFFF;[email protected];[email protected][email protected]@[email protected];@
      @M02846:68:000000000-C39D4:1:1101:23101:3568 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected][email protected]@@@@@@@@@@@@
      @M02846:68:000000000-C39D4:1:1101:8961:3608 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >AAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFAFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF
      @M02846:68:000000000-C39D4:1:1101:8961:3608 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@
      @M02846:68:000000000-C39D4:1:1101:8333:3768 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>AFGAEGECECECE/>[email protected]@@@[email protected]@@<@@;@@>>@@[email protected]@@@@@[email protected]@[email protected]@@[email protected][email protected]@@;@@@;@@@;?--9>;@[email protected]@->@@[email protected]@[email protected]@@@@;@@[email protected][email protected]@@;-->@@@@@[email protected][email protected]=
      @M02846:68:000000000-C39D4:1:1101:8333:3768 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@<@@@[email protected]@@@@@@@@@@?
      @M02846:68:000000000-C39D4:1:1101:17863:3914 1:N:0:1


      Thanks a lot to all,

      Nicolas
      Nicolas Tremblay
      Graduate Student

      Cardiovascular Genetics - Andelfinger Lab
      CHU Ste-Justine Research Center

      Comment


      • #4
        Code:
        @m02846:68:000000000-c39d4:1:1101:14422:2079 1:n:0:1
        tttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttt
        +
        >>afggggegggggggggggggggggggggggfggggggggggggggfffffffffffffffffffffffffffffffffffffffffffffffffffabffffffffffffffffffffffffffffffffffffffffff
        @m02846:68:000000000-c39d4:1:1101:14422:2079 2:n:0:1
        aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa
        +
        [email protected][email protected]@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@@@[email protected][email protected]<@@@@@@@@[email protected][email protected]@[email protected]@@@@@@@@@@[email protected]@@@@[email protected]@;;;=?;[email protected]@@[email protected]
        @m02846:68:000000000-c39d4:1:1101:14218:2110 1:n:0:1
        tttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttt
        +
        >>[email protected]@@/<[email protected]@cdgcbf?9bdab;dffffffbffffba=affffaffbbfbfffaafffaf;fffdffffffbd=ddf-9>[email protected][email protected]@bdf;[email protected];9=
        M02846:68:000000000-C39D4:1:1101:14218:2110 2:N:0:1
        AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
        +
        [email protected]@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@;[email protected]>[email protected]@;@
        @M02846:68:000000000-C39D4:1:1101:13886:2230 1:N:0:1
        TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
        +
        >>AGGGGGGGGEEFGGGGFE/@[email protected][email protected]>DFFFAA;@[email protected]=FBFAABCF;BDDDC-B;-B;FD;>[email protected]@[email protected][email protected]
        @M02846:68:000000000-C39D4:1:1101:13886:2230 2:N:0:1
        AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
        +
        [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@@[email protected]@@[email protected]@@@
        @M02846:68:000000000-C39D4:1:1101:12194:2260 1:N:0:1
        TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
        +
        >>AFGGGGGGGGGGGCGGGGGGGGGGGGGGGGGGGFFGFGGGGGGGGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFAAFFFFFA=DFFFA;FF;AFFFFFFFFFFFAFF;;ADAFFAFCFFFFAC;C
        @M02846:68:000000000-C39D4:1:1101:12194:2260 2:N:0:1
        AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
        +
        [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@
        @M02846:68:000000000-C39D4:1:1101:12722:2310 1:N:0:1
        TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
        +
        >>[email protected]/[email protected]@@[email protected]?BBBBB;@B?;[email protected][email protected]?B?B?B;>[email protected]>;-;BBB99B-B;[email protected];[email protected];=>@;[email protected]
        @M02846:68:000000000-C39D4:1:1101:12722:2310 2:N:0:1
        AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
        +
        [email protected]@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@[email protected][email protected]@@@@@@@@[email protected]@@@[email protected]@@@@@@@@@@@@@@@@@[email protected]@@@[email protected]@@@@@@[email protected]@@
        @M02846:68:000000000-C39D4:1:1101:20711:2313 1:N:0:1
        TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
        +
        >>AGGGGGGGGGGGGGGGGGGGCFGFGGGGCCGGGGGGGGGGGGGGGFAFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFBFFFFFFFFCAFFFFFFCFFFFFFFFFFFFFFFFFFAFFFFFAFFFFFF
        @M02846:68:000000000-C39D4:1:1101:20711:2313 2:N:0:1
        AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
        +
        [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@
        @M02846:68:000000000-C39D4:1:1101:17123:2486 1:N:0:1
        TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTATTGTTTTGGTGGGGTTTTTTGTGACTTCGACACTGTTT
        +
        >BBGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFDFA=D----.0000://.-<..--.-...--</00000....-:0000
        @M02846:68:000000000-C39D4:1:1101:17123:2486 2:N:0:1
        AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
        +
        [email protected]@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@<[email protected]@@[email protected]@[email protected]@[email protected][email protected]@[email protected];[email protected]@;=->[email protected]?;[email protected]<=;[email protected]@-
        @M02846:68:000000000-C39D4:1:1101:19948:2714 1:N:0:1
        TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTATAATTTTAATATTTTCGGGTGGGCTATACAACTGCCTTTTTCCTTCTTTTCCT
        +
        >>[email protected]@@GG<FFFFFFFFFFFFFFFFFFFFFFFDFFFFFAFFFFAF---9.////9///////;///--..---.;.///////;/////;/9/://;/////;
        @M02846:68:000000000-C39D4:1:1101:19948:2714 2:N:0:1
        AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAACAAAAAAACAAAAAAAAAAAAAAAAAAAAAAATAAAAAAAAACAAACC
        +
        [email protected]@@@@@@@@@@@[email protected]@@@@@@@@@[email protected][email protected]@@@[email protected][email protected]@[email protected]?<@@--9A--;@-9-;A-=---9-99;@---9-9-;-;////----------
        @M02846:68:000000000-C39D4:1:1101:14946:3025 1:N:0:1
        TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
        +
        >AAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF
        @M02846:68:000000000-C39D4:1:1101:14946:3025 2:N:0:1
        AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
        +
        [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
        @M02846:68:000000000-C39D4:1:1101:15839:3063 1:N:0:1
        TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
        +
        3>>[email protected]@BBBB??B<BBBBBBB?;[email protected]?B<[email protected];BB?BB;B??BB?>=?;[email protected]@??BBBBB?--BB?-BBB?
        @M02846:68:000000000-C39D4:1:1101:15839:3063 2:N:0:1
        AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
        +
        [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@[email protected]@@@@@@@
        @M02846:68:000000000-C39D4:1:1101:10519:3107 1:N:0:1
        TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
        +
        >>AGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF;BFFFFFFFFAFAFFFF
        @M02846:68:000000000-C39D4:1:1101:10519:3107 2:N:0:1
        AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
        +
        [email protected]@@@@@[email protected]@@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@[email protected]@@@@@@@@@@@@@@@@[email protected]@@@@@@[email protected][email protected]@@@@[email protected]@@@
        @M02846:68:000000000-C39D4:1:1101:14081:3115 1:N:0:1
        TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
        +
        >>AGGGGGGGEGGGGGCGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGFAFFFFADFFFFFFFFFFFFFFFFFAFFFAFFFFFFFFFABFAFAFFFFFAF;DFFFFFFFAFAFFFAFFFAFFFFFFFFFAFFAFFAACFFACC
        @M02846:68:000000000-C39D4:1:1101:14081:3115 2:N:0:1
        AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
        +
        [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
        @M02846:68:000000000-C39D4:1:1101:10255:3290 1:N:0:1
        TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
        +
        >>[email protected]FFFFFFFFFFFFFFFFAFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFAFAFFFACFFFFFFFFFAFFF
        @M02846:68:000000000-C39D4:1:1101:10255:3290 2:N:0:1
        AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
        +
        [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
        @M02846:68:000000000-C39D4:1:1101:20826:3326 1:N:0:1
        TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
        +
        >AAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFF
        @M02846:68:000000000-C39D4:1:1101:20826:3326 2:N:0:1
        AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
        +
        [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
        @M02846:68:000000000-C39D4:1:1101:9032:3354 1:N:0:1
        TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
        +
        >>AGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFAFFFFFFFFAFFFFFFFAFFFFFFFFFFFFFFFFCAFF
        @M02846:68:000000000-C39D4:1:1101:9032:3354 2:N:0:1
        AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
        +
        [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
        @M02846:68:000000000-C39D4:1:1101:21178:3402 1:N:0:1
        TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
        +
        >>AFGFGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGFFFFFFFFFFFFFFFFFFFFFFFAFFFFFFFFFFFFFFFFFFFFAAFFFFFFFFFFF;DACFFFFFFFFFFF-DFFFFAAFFAFFFFFFFFFBFF
        @M02846:68:000000000-C39D4:1:1101:21178:3402 2:N:0:1
        AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
        +
        [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@@@@@@@@@@@@@@
        @M02846:68:000000000-C39D4:1:1101:18043:3408 1:N:0:1
        TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
        +
        3>AEGGGGGGGGGGGGGGGGGGGCGGGGGGGGGGGGGGGGGGGGGGGFFFFFFFFFFF-BBFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFAFCFAA;BFFFFFFFFFFFFFFFFFFFFFFAAB;@B--;--;;-;
        @M02846:68:000000000-C39D4:1:1101:18043:3408 2:N:0:1
        AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
        +
        [email protected]@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@[email protected]@@[email protected]@@@@@[email protected][email protected]@@@@@@@@@
        @M02846:68:000000000-C39D4:1:1101:12925:3455 1:N:0:1
        TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTGTATACTTAATTTCTTTGAGTGTCTGAGTAATGTCTGACACAGCCACCTTATTATT
        +
        >>[email protected]@[email protected]@GBBBBBB?BBBBB??<[email protected]?BBBBB?BB?B;B=BB;B->-9/////////////////.;///9//;///////////9/...9....///;9/9
        Last edited by GenoMax; 10-16-2018, 08:57 AM. Reason: Added [code] tags
        Nicolas Tremblay
        Graduate Student

        Cardiovascular Genetics - Andelfinger Lab
        CHU Ste-Justine Research Center

        Comment


        • #5
          I do not know how the library has been prepped but I notice following:

          1- It is stranded (directional) library
          2- Probably RNA has been poly adenylated after extraction
          3- It contains some short fragments (below 100 base)
          4- RT reaction probably has been primed with polyT primer

          It is library prep issue and not the sequencing. If you would like help you need to give more info on extraction and library prep.

          Comment


          • #6
            Viral RNA was extracted from plasma samples using QIAamp Viral RNA Mini Kit (720uL of sample) using standard protocol.

            Double stranded cDNA synthesis was done using Maxima H Minus Double-Stranded cDNA Synthesis Kit (random hexamers, Oligo (dT) and HCV specific pan genomic primers dA20) following standard protocol cycling conditions.

            The dsCDNA was quantified by nanodrop and promega QuantiFluor® dsDNA System.

            I will inquired how the cDNA was prep for sequencing.

            Thanks,

            Nicolas
            Nicolas Tremblay
            Graduate Student

            Cardiovascular Genetics - Andelfinger Lab
            CHU Ste-Justine Research Center

            Comment


            • #7
              I do not know why oligo dT has been used in 1st strand synthesis. It is possible that dA20 oligo has hybridized to oligo T resulting in creating long stretches of T by joining that has been used as template by reverse transcriptase. Because of high primer concentration polyT artefact has been dominant species in the reactions and has ended up in final library.

              Comment

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