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  • JonB
    Member
    • Jan 2010
    • 85

    #1

    SmallRNA: how to trim poly-a stretch + other sequence at 5'-end?

    Hi,

    I have sequenced smallRNA sequences on Illumina HiSeq single end.
    My reads are 100 nts, and at the 3'-end is the poly-a stretch from the oligo-dT adapter and sometimes also other nucleotides when the oligo-dT was sequenced through. But I do not understand where these come from (does not match the adapter it seems). Can someone suggest a good way to trim these reads from the poly-A stretch?

    Sample seqs:

    Code:
    @HWI-ST486:386:D1UMHACXX:3:1101:1399:2119 1:N:0:TGACCA
    NTCGTATCTGTAATCATGAACTTGTCAACGGCTACCTGGTTTCTGTCCTAAAAAAAAAAAAAAAAAAAAAAAAAAAAGGGAAGACCAAACTCTGGAACCCC
    +
    #1:BDBEDFHHGHGCBGHGCHHIJGIFGGIGEHCHJJIJHGH<FDGDHF@D@ACFEH6A>?BBB@>>B>BBB#############################
    @HWI-ST486:386:D1UMHACXX:3:1101:1367:2139 1:N:0:TGACCA
    NATGTTTTTTGGGGTTATAACAGGGTGGAGCGCTTTATGCGACTTCGCCCTTTAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
    +
    #1=DDDEDHHDHHJJIGIFDFGIIJHGI?GHJIFHI@GG@F@;AH=?=BDCEE@@>@ABBDBBBD;BBBDDDDBB@BBDBBD@DBB>B@>>>5-59@<@BD
    @HWI-ST486:386:D1UMHACXX:3:1101:1392:2197 1:N:0:TGACCA
    ACAATTTATGGTAGAGAAAAAAAAAAAAAAAAAAAAAAAAAAAGGGGAAGAGCAAAGGTCTGAAACCCAGCCATTTACCATTCTCCTTTTCCCCTTTTTTT
    +
    CCCFFFFFHHHFFIIIIIIIIIFFGGGBBEDCBB=@#################################################################
    Adapters used:
    Code:
    TrueSeq_Sense_primer
    5'-AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACGCTCTTCCGATCT-3’
    TrueSeq_Antisense_ NNNNNN_primer Barcode
    5’-CAAGCAGAAGACGGCATACGAGAT-NNNNNN-GTGACTGGAGTTCAGACGTGTGCTCTTCCGATC(dT)25-3’
    Thanks!

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