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  • #1

    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
    Tags: None
  • #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
      +
      BFFCCECCEGGGGGGGGGGGGGGCEGGGGGGGGGGGGGGGGGGGGGGGG@?@@@@@@@@@@@@@@@?@@@@@@@@@@@@@@@@@@@?@??@<@@@@@@@@?@?@@??@@@@@@@@@@@???@@@@@?@@;;;=?;=@@@??@
      @M02846:68:000000000-C39D4:1:1101:14218:2110 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>AGGGGFEFGCFEGGG?EE@EGGGEGCC@@/<AFC@@CDGCBF?9BDAB;DFFFFFFBFFFFBA=AFFFFAFFBBFBFFFAAFFFAF;FFFDFFFFFFBD=DDF-9>99B=9@==9-9@-9@BDF;BCFF@DCBFFA-;9=
      @M02846:68:000000000-C39D4:1:1101:14218:2110 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      BFAGGCEEEEGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@?@@@@@@@@@@@@@@@@@@@@@@??@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@@@@;--@->-@99@;@
      @M02846:68:000000000-C39D4:1:1101:13886:2230 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>AGGGGGGGGEEFGGGGFE/@EEGGCCCDGGG?@DCGFFGGB??DGFFFFFF@D>DFFFAA;@FFDABFFAFA@DADDFFFFFFFFA=FBFAABCF;BDDDC-B;-B;FD;>-@FCAFAD--9@BDFAFFFFAFC-@D=@D
      @M02846:68:000000000-C39D4:1:1101:13886:2230 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      DFFGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@@@@@@@@@@@@@@@@@@@?@@@?@@@@
      @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
      +
      CFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@@@@@@@@@@@@@
      @M02846:68:000000000-C39D4:1:1101:12722:2310 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>AFGEEGAEGE0EFCEEEE@EG/EECCGGGCCCCCC@DCC@@BB@B?BBBBB;@B?;BB==BBBBBBBB?BBBBBBBB?B-9@B?B?@BBB?B?B?B;>BB@B>;-;BBB99B-B;BB--9@-99-B-;B--@;=>@;--@
      @M02846:68:000000000-C39D4:1:1101:12722:2310 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      BFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@?@@@@@@@@@@@@@@@@@@=@@@@@@@@@@@?@?@@@@@@@@@??@@@@?@@@@@@@@@@@@@@@@@@?@@@@?@@@@@@@?@@@
      @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
      +
      CFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@
      @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
      +
      BFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@?@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@<=@@@?@@?@@?@?@@?@;??-9@@;=->=@?;?=@<=;-9=@@-
      @M02846:68:000000000-C39D4:1:1101:19948:2714 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTATAATTTTAATATTTTCGGGTGGGCTATACAACTGCCTTTTTCCTTCTTTTCCT
      +
      >>AFFGGGGGCGGGG?GCEFGFGGGGGGGFGGGGGGGGGGCG@@@GG<FFFFFFFFFFFFFFFFFFFFFFFDFFFFFAFFFFAF---9.////9///////;///--..---.;.///////;/////;/9/://;/////;
      @M02846:68:000000000-C39D4:1:1101:19948:2714 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAACAAAAAAACAAAAAAAAAAAAAAAAAAAAAAATAAAAAAAAACAAACC
      +
      FFFGFGGGGGGGGGGGGEGGGGGGGGGGGGGGGGCGGGGGGGGGGAGAG@@@@@@@@@@@@?@@@@@@@@@@??@?@@@@=?@?@-@-9@?<@@--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
      +
      CFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
      @M02846:68:000000000-C39D4:1:1101:15839:3063 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      3>>FFGCEGGGGEGG?EEGCGCGGEECC?C@CCGGGGGGGGGGGGG@BBBB??B<BBBBBBB?;?BBBB?BBBBBB?BB-@BBBBB?B<BBBBBBBB-@;BB?BB;B??BB?>=?;B@B-BB-@??BBBBB?--BB?-BBB?
      @M02846:68:000000000-C39D4:1:1101:15839:3063 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      BFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@?@@@@@@@@
      @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
      +
      BFFGGGEGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@?@@@@@@@?@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@@?@@@@@@@@@@@@@@@@@?@@@@@@@?@?@@@@@?@@@@
      @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
      +
      BFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
      @M02846:68:000000000-C39D4:1:1101:10255:3290 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>AFFGGFGGGFGGFGGGGCGGGGGGGGGGGGGGGGGGGGCG@GGGGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFAFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFAFAFFFACFFFFFFFFFAFFF
      @M02846:68:000000000-C39D4:1:1101:10255:3290 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      CFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
      @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
      +
      CFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
      @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
      +
      CFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
      @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
      +
      BFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@@@@@@@@@@@@
      @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
      +
      BFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@?@@@@@@?@?@@@@@@@@@@
      @M02846:68:000000000-C39D4:1:1101:12925:3455 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTGTATACTTAATTTCTTTGAGTGTCTGAGTAATGTCTGACACAGCCACCTTATTATT
      +
      >>AFFEEEGGGGGGGGEGEEGGCGGCCGC@C@CCCGC@DCCDGGG@GBBBBBB?BBBBB??<BB-@?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
      +
      DFFGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@?@@@@<@@@?@@@@@@?@@@@@@<@?@@?;=@?@@??@;@@?@-@----------9---9-;--;------------9--//
      @M02846:68:000000000-C39D4:1:1101:23101:3568 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>AFFEGGGCGG?EGGGGGGGEG?EGGGGCGC?D@CCGCCGGGGGGGFF=BFFF-DFBFAFFFFFFFFBFADFFFFFFFDAFFFFFFFF;D9D-@FFFFFFFADDADF;-99@DFFF=@D-B@BB=B9@DFFFFFFFFFA;@
      @M02846:68:000000000-C39D4:1:1101:23101:3568 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      CFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@?@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@?@@@@@@@@@@@@@
      @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
      +
      BFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@?@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
      @M02846:68:000000000-C39D4:1:1101:8333:3768 1:N:0:1
      TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
      +
      >>AFGAEGECECECE/>FEEGGGGGGGGGG@CC@@@C?CCGCGGGGG@@@<@@;@@>>@@?@@@@@@?@@?@@@?@?@@@;@@@;@@@;?--9>;@--9-@@->@@?-@@?@@@@@;@@?@???@-@@;-->@@@@@?@?@=
      @M02846:68:000000000-C39D4:1:1101:8333:3768 2:N:0:1
      AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
      +
      BFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@@@@@@@@@@@@<@@@??@@@@@@@@@@@?
      @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
+
bffccecceggggggggggggggcegggggggggggggggggggggggg@?@@@@@@@@@@@@@@@?@@@@@@@@@@@@@@@@@@@?@??@<@@@@@@@@?@?@@??@@@@@@@@@@@???@@@@@?@@;;;=?;=@@@??@
@m02846:68:000000000-c39d4:1:1101:14218:2110 1:n:0:1
tttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttttt
+
>>aggggfefgcfeggg?ee@egggegcc@@/<afc@@cdgcbf?9bdab;dffffffbffffba=affffaffbbfbfffaafffaf;fffdffffffbd=ddf-9>99b=9@==9-9@-9@bdf;bcff@dcbffa-;9=
M02846:68:000000000-C39D4:1:1101:14218:2110 2:N:0:1
AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
+
BFAGGCEEEEGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@?@@@@@@@@@@@@@@@@@@@@@@??@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@@@@;--@->-@99@;@
@M02846:68:000000000-C39D4:1:1101:13886:2230 1:N:0:1
TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
+
>>AGGGGGGGGEEFGGGGFE/@EEGGCCCDGGG?@DCGFFGGB??DGFFFFFF@D>DFFFAA;@FFDABFFAFA@DADDFFFFFFFFA=FBFAABCF;BDDDC-B;-B;FD;>-@FCAFAD--9@BDFAFFFFAFC-@D=@D
@M02846:68:000000000-C39D4:1:1101:13886:2230 2:N:0:1
AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
+
DFFGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@@@@@@@@@@@@@@@@@@@?@@@?@@@@
@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
+
CFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@@@@@@@@@@@@@
@M02846:68:000000000-C39D4:1:1101:12722:2310 1:N:0:1
TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
+
>>AFGEEGAEGE0EFCEEEE@EG/EECCGGGCCCCCC@DCC@@BB@B?BBBBB;@B?;BB==BBBBBBBB?BBBBBBBB?B-9@B?B?@BBB?B?B?B;>BB@B>;-;BBB99B-B;BB--9@-99-B-;B--@;=>@;--@
@M02846:68:000000000-C39D4:1:1101:12722:2310 2:N:0:1
AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
+
BFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@?@@@@@@@@@@@@@@@@@@=@@@@@@@@@@@?@?@@@@@@@@@??@@@@?@@@@@@@@@@@@@@@@@@?@@@@?@@@@@@@?@@@
@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
+
CFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@
@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
+
BFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@?@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@<=@@@?@@?@@?@?@@?@;??-9@@;=->=@?;?=@<=;-9=@@-
@M02846:68:000000000-C39D4:1:1101:19948:2714 1:N:0:1
TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTATAATTTTAATATTTTCGGGTGGGCTATACAACTGCCTTTTTCCTTCTTTTCCT
+
>>AFFGGGGGCGGGG?GCEFGFGGGGGGGFGGGGGGGGGGCG@@@GG<FFFFFFFFFFFFFFFFFFFFFFFDFFFFFAFFFFAF---9.////9///////;///--..---.;.///////;/////;/9/://;/////;
@M02846:68:000000000-C39D4:1:1101:19948:2714 2:N:0:1
AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAACAAAAAAACAAAAAAAAAAAAAAAAAAAAAAATAAAAAAAAACAAACC
+
FFFGFGGGGGGGGGGGGEGGGGGGGGGGGGGGGGCGGGGGGGGGGAGAG@@@@@@@@@@@@?@@@@@@@@@@??@?@@@@=?@?@-@-9@?<@@--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
+
CFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
@M02846:68:000000000-C39D4:1:1101:15839:3063 1:N:0:1
TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
+
3>>FFGCEGGGGEGG?EEGCGCGGEECC?C@CCGGGGGGGGGGGGG@BBBB??B<BBBBBBB?;?BBBB?BBBBBB?BB-@BBBBB?B<BBBBBBBB-@;BB?BB;B??BB?>=?;B@B-BB-@??BBBBB?--BB?-BBB?
@M02846:68:000000000-C39D4:1:1101:15839:3063 2:N:0:1
AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
+
BFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@?@@@@@@@@
@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
+
BFFGGGEGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@?@@@@@@@?@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@@?@@@@@@@@@@@@@@@@@?@@@@@@@?@?@@@@@?@@@@
@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
+
BFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
@M02846:68:000000000-C39D4:1:1101:10255:3290 1:N:0:1
TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
+
>>AFFGGFGGGFGGFGGGGCGGGGGGGGGGGGGGGGGGGGCG@GGGGFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFAFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFFAFAFFFACFFFFFFFFFAFFF
@M02846:68:000000000-C39D4:1:1101:10255:3290 2:N:0:1
AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
+
CFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
@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
+
CFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
@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
+
CFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@
@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
+
BFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@@@@@@@@@@@@@@
@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
+
BFAGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGGG@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@@?@@@?@@@@@@?@?@@@@@@@@@@
@M02846:68:000000000-C39D4:1:1101:12925:3455 1:N:0:1
TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTGTATACTTAATTTCTTTGAGTGTCTGAGTAATGTCTGACACAGCCACCTTATTATT
+
>>AFFEEEGGGGGGGGEGEEGGCGGCCGC@C@CCCGC@DCCDGGG@GBBBBBB?BBBBB??<BB-@?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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