Hello
I have a weird 454 result produced by MID and DSN normalization.
The mRNA reads started with 'key(4bp)+MID(10bp)+cDNA library primer (20~25bp) sequence from 5' end. The reads are supposed to included key+MID, not cDNA library primer. I feel like that there seemed to be something wrong with cDNA library construction. Below is the sample reads.
Are they normal?
>GRMETAB01A1BYK
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTGGGAGAATCTCGAACCACATCTCTGCCCGTACCTAGCaacctaactctggtcgtnaagagctgntcncctcccncctcctntgtgtgtactactacgctcttntagacagcacgacgacacacggaggtaggagnn
>GRMETAB01AK1L7
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTGACTGAAACACAATTCATTTCTGACTGACATTacctttctgtcaactagacggtggcaccacaaataataccggtctttgtgcttgtctcttctagatgganggtttgttaaggactcttgttgtcattactttaaaaagaagtaataaaggattttaaagtaacgaaaaaacgaaaagaaaaactaaaactaaacgtcgtgtagtacactacacgtctctgagagcacgacgacacacggaggtaggngnnnnn
>GRMETAB01BC7E6
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTACGGAGGTGTTGAAGaagattgaatcaagaactaacagccaaagaagaagaagaagaagaagacagaagaggagcagaagaaactaagacacagattcaagaatctcaggtaacatccaccaacaaggaacggacggttcctaacgacgggagtgagaactaagacaagaaatactcaagaccactggaggaggctaaagacagagcgagaaaacgaaagaacagaacaagctgagctgagactgccaacggacacacagggagtaggnnnn
>GRMETAB01EOICN
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTACTGAGGGAAGTAGTTGTGTTGGCTGAGTTGTCAGTCAAGCGAATTCCGCTACTCTGCGTTGATACCACTGCTctgagactgccaaggcacacagggataggnnnnnnnnnn
>GRMETAB01DR31L
tcagACGAGTGCGTAGCAGTGGTATCAACGCAGAGTACTTTTTTTTTNTTTNTTTAttataactaactaactatatatattattattatatatatctctactactactactactatatactactatatatatactactactacaccacacacgtagtagtagtacgtactactactactacgagagagagagagagacgagtaggtagtacgtacgtacgtacgacgacgtactactaactaactaactacgtacgacgacgacgacgacgacgacgtacgtacgtaccgtaccgtaccgtaccgtaccgtaccgacctacctaacctacctaacctactactactacgtacgacgacgacgacgacgacgacgagagagngnnnn
>GRMETAB01A4CCL
tcagACGAGTGCGTAGCAGTGGTATCAACGCAGAGTTACTTTTTTTTTTTTTTTTTTTTTNttttnttttnttttnttttnttttntttatttatttatttatttatttatttattattattatatatatatatatatatatatatatatatatatatagtatatatatatatatatatatatatatatatatatatatatagtatagtagtagtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgacgacgacgacgacgacgagagagagagngnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn
>GRMETAB01BL4UE
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTACTTTNTTTNTTTCTTTCTTTCTTTCTTTCttctctactatctacaanagacacacactctctctcgtctcgtcgtacgtacgtacgtcgacgacgacgacgacgacgacgagagagngnnn
>GRMETAB01BWBZ9
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGACAGCCAAGTCATTGCCAGACACATACAAAGATAAGCCCACGTActtacngcgttgataccacgtcggcttctgaggacgttggaccaaagggacgacctaacctagggggaattaggggngcncncnngagagaggngcngtnttnanntgngnngagangcacagatacgctcgcagacgtgggttcggtccgacgacccgtaacggtaacctacaacgcttcctttaggttatagtaattaaatttagtacttatttcgttcggacggacgaacgaaaaacgggaaaccgtaacgtaccaccacgtacctactaggacaaacaactacttcttcgtcgtctagtaggacgagggaggacgtacgtaggtagtttattattaattacttctagaagacacagagacgacgaccaacgtactattcgaccgaacgaacaacaactaaacaaaacgaaacgtaaaccgacccgtaacgtacttacgtcgtcgttacgtaactaacctactcgtcttcgtacgacgacgtacgaccgaacggacgacgaacggaagggaaggtaggagnnnnn
>GRMETAB01C584Q
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTACAGGGGTGTTTACAAGAGATGGTTTGCAATGCAGACTaagcagaaaatttagtagngtgatggataggaaaccaaaccaacagcccccgtacctctncgtgtatacacctgctcttcgagactgcacggacgacacaggggagtagngnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn
>GRMETAB01AOKGH
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTACGTTTNTTTNTTTTTTTTTNTTTTTTNTTTTTNttttnttttntttttattttattttattttattttattttattttatttatttatttatttatttatttatttatttattacttacttattagtagtagtagtagtacgtagtacgtacgtacgtacgtacgtacgtacgtacgtactacgtacgtacgtacgtacgtacgtactactactactactactactactacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgacgacgacgacgacgacgacgagacgacgagagannnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn
>GRMETAB01D7O01
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTACGGGAATTATGACATGCAccgatttacatcccncctctcngtcgtatanacctatctccttcagaagtnttctacgagaaccaacctaggnggacgagatntctcntacgtcttgtacganctacctcgngnnntcncntctcgttgtggtggttcttacgtcgttcgttgtagttgtaagtaacgtacgtgagagaagacgtacgtacgactacctacgtacgtacaactaaagttaaagttaaattaaaaagtaaaaagttaaagtttaagtttaccgttaccgtacgtaggtaaggaaaggaaaggaacgaacgtaaacgtttaaccgtttaaaccgtaacgaaccgaacccgaaaccgtaaacggtaaaaacggaaaaaccgaaaaccgaaacgtaagtacgaacggaaacggtaacgtaacggaaggaaggaacgaaaccgtaaaacgttaaacgttaaacgtaaacgtaaaacgtaaacgtaaacgtaacctaaccgtaacgtaagttacgtacgtagtagtaacgtaacgaaacgaaccgtaaccgtaaacgtaaactaacgtaacgtaaccgttaccgttaccgttacccgttaacccttaaaccccttaacccttacgttacgtaacggtaacggtaaaggttaaagttaagtaacgtaacgtacgtacgtaaccgtaaccgttaaccgttaaccttaaccccgttaccccgttacccgttaccgttacggttacgttacgtaaccgtaaccgtaccgttaccgttaccgttacgtacgtacg
>GRMETAB01AHC9E
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTAGGTGATGGAAACCTTTGGTAGACTCTTCTAAGTGCACGTGCAGATTTTCTCTTACCATGTATTTTACTTAAACAAATTTGTTTGTTATTTagtccacccgcctccttcttctacacacctctcgttccttcctaccgccactacttaacttagttgtcggttccccgttaccttcctaccgtccgttntgtcgtactanaccttcctgttgaccttgacgaagccacgaccgaagggactaggaggaggnngnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn
I have a weird 454 result produced by MID and DSN normalization.
The mRNA reads started with 'key(4bp)+MID(10bp)+cDNA library primer (20~25bp) sequence from 5' end. The reads are supposed to included key+MID, not cDNA library primer. I feel like that there seemed to be something wrong with cDNA library construction. Below is the sample reads.
Are they normal?
>GRMETAB01A1BYK
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTGGGAGAATCTCGAACCACATCTCTGCCCGTACCTAGCaacctaactctggtcgtnaagagctgntcncctcccncctcctntgtgtgtactactacgctcttntagacagcacgacgacacacggaggtaggagnn
>GRMETAB01AK1L7
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTGACTGAAACACAATTCATTTCTGACTGACATTacctttctgtcaactagacggtggcaccacaaataataccggtctttgtgcttgtctcttctagatgganggtttgttaaggactcttgttgtcattactttaaaaagaagtaataaaggattttaaagtaacgaaaaaacgaaaagaaaaactaaaactaaacgtcgtgtagtacactacacgtctctgagagcacgacgacacacggaggtaggngnnnnn
>GRMETAB01BC7E6
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTACGGAGGTGTTGAAGaagattgaatcaagaactaacagccaaagaagaagaagaagaagaagacagaagaggagcagaagaaactaagacacagattcaagaatctcaggtaacatccaccaacaaggaacggacggttcctaacgacgggagtgagaactaagacaagaaatactcaagaccactggaggaggctaaagacagagcgagaaaacgaaagaacagaacaagctgagctgagactgccaacggacacacagggagtaggnnnn
>GRMETAB01EOICN
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTACTGAGGGAAGTAGTTGTGTTGGCTGAGTTGTCAGTCAAGCGAATTCCGCTACTCTGCGTTGATACCACTGCTctgagactgccaaggcacacagggataggnnnnnnnnnn
>GRMETAB01DR31L
tcagACGAGTGCGTAGCAGTGGTATCAACGCAGAGTACTTTTTTTTTNTTTNTTTAttataactaactaactatatatattattattatatatatctctactactactactactatatactactatatatatactactactacaccacacacgtagtagtagtacgtactactactactacgagagagagagagagacgagtaggtagtacgtacgtacgtacgacgacgtactactaactaactaactacgtacgacgacgacgacgacgacgacgtacgtacgtaccgtaccgtaccgtaccgtaccgtaccgacctacctaacctacctaacctactactactacgtacgacgacgacgacgacgacgacgagagagngnnnn
>GRMETAB01A4CCL
tcagACGAGTGCGTAGCAGTGGTATCAACGCAGAGTTACTTTTTTTTTTTTTTTTTTTTTNttttnttttnttttnttttnttttntttatttatttatttatttatttatttattattattatatatatatatatatatatatatatatatatatatagtatatatatatatatatatatatatatatatatatatatatagtatagtagtagtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgacgacgacgacgacgacgagagagagagngnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn
>GRMETAB01BL4UE
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTACTTTNTTTNTTTCTTTCTTTCTTTCTTTCttctctactatctacaanagacacacactctctctcgtctcgtcgtacgtacgtacgtcgacgacgacgacgacgacgacgagagagngnnn
>GRMETAB01BWBZ9
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGACAGCCAAGTCATTGCCAGACACATACAAAGATAAGCCCACGTActtacngcgttgataccacgtcggcttctgaggacgttggaccaaagggacgacctaacctagggggaattaggggngcncncnngagagaggngcngtnttnanntgngnngagangcacagatacgctcgcagacgtgggttcggtccgacgacccgtaacggtaacctacaacgcttcctttaggttatagtaattaaatttagtacttatttcgttcggacggacgaacgaaaaacgggaaaccgtaacgtaccaccacgtacctactaggacaaacaactacttcttcgtcgtctagtaggacgagggaggacgtacgtaggtagtttattattaattacttctagaagacacagagacgacgaccaacgtactattcgaccgaacgaacaacaactaaacaaaacgaaacgtaaaccgacccgtaacgtacttacgtcgtcgttacgtaactaacctactcgtcttcgtacgacgacgtacgaccgaacggacgacgaacggaagggaaggtaggagnnnnn
>GRMETAB01C584Q
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTACAGGGGTGTTTACAAGAGATGGTTTGCAATGCAGACTaagcagaaaatttagtagngtgatggataggaaaccaaaccaacagcccccgtacctctncgtgtatacacctgctcttcgagactgcacggacgacacaggggagtagngnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn
>GRMETAB01AOKGH
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTACGTTTNTTTNTTTTTTTTTNTTTTTTNTTTTTNttttnttttntttttattttattttattttattttattttattttatttatttatttatttatttatttatttatttattacttacttattagtagtagtagtagtacgtagtacgtacgtacgtacgtacgtacgtacgtacgtactacgtacgtacgtacgtacgtacgtactactactactactactactactacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgtacgacgacgacgacgacgacgacgagacgacgagagannnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn
>GRMETAB01D7O01
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTACGGGAATTATGACATGCAccgatttacatcccncctctcngtcgtatanacctatctccttcagaagtnttctacgagaaccaacctaggnggacgagatntctcntacgtcttgtacganctacctcgngnnntcncntctcgttgtggtggttcttacgtcgttcgttgtagttgtaagtaacgtacgtgagagaagacgtacgtacgactacctacgtacgtacaactaaagttaaagttaaattaaaaagtaaaaagttaaagtttaagtttaccgttaccgtacgtaggtaaggaaaggaaaggaacgaacgtaaacgtttaaccgtttaaaccgtaacgaaccgaacccgaaaccgtaaacggtaaaaacggaaaaaccgaaaaccgaaacgtaagtacgaacggaaacggtaacgtaacggaaggaaggaacgaaaccgtaaaacgttaaacgttaaacgtaaacgtaaaacgtaaacgtaaacgtaacctaaccgtaacgtaagttacgtacgtagtagtaacgtaacgaaacgaaccgtaaccgtaaacgtaaactaacgtaacgtaaccgttaccgttaccgttacccgttaacccttaaaccccttaacccttacgttacgtaacggtaacggtaaaggttaaagttaagtaacgtaacgtacgtacgtaaccgtaaccgttaaccgttaaccttaaccccgttaccccgttacccgttaccgttacggttacgttacgtaaccgtaaccgtaccgttaccgttaccgttacgtacgtacg
>GRMETAB01AHC9E
tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTAGGTGATGGAAACCTTTGGTAGACTCTTCTAAGTGCACGTGCAGATTTTCTCTTACCATGTATTTTACTTAAACAAATTTGTTTGTTATTTagtccacccgcctccttcttctacacacctctcgttccttcctaccgccactacttaacttagttgtcggttccccgttaccttcctaccgtccgttntgtcgtactanaccttcctgttgaccttgacgaagccacgaccgaagggactaggaggaggnngnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn
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