Seqanswers Leaderboard Ad

Collapse

Announcement

Collapse
No announcement yet.
X
 
  • Filter
  • Time
  • Show
Clear All
new posts

  • weird 454 reads

    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

  • #2
    TTTNTTTNTTTTTTTTTNTTTTTTNTTTTTN looks like a poly A primer, broken up to avoid a huge homopolymer run.

    I'll also bet that the >GRMETAB01A4CCL read hit that wall of T's and died immediately, hence 'tatata'.

    Comment


    • #3
      Originally posted by GW_OK View Post
      TTTNTTTNTTTTTTTTTNTTTTTTNTTTTTN looks like a poly A primer, broken up to avoid a huge homopolymer run.

      I'll also bet that the >GRMETAB01A4CCL read hit that wall of T's and died immediately, hence 'tatata'.
      I still wondering whether the reads including poly-A primer are common or not. I mean which one is normal read?

      (1)
      tcagACGAGTGCGTAAGCAGTGGTATCAACGCAGAGTGGGAGAATCTCGAACCACATCTCTGCCCGTACCTAGCaacctaactctggtcgtnaagagctgntcncctcccncctcctntgtgtgtactactacgctcttntagacagcacgacgacacacggaggtaggagnn

      (2)
      tcagACGAGTGCGTGGGAGAATCTCGAACCACATCTCTGCCCGTACCTAGCaacctaactctggtcgtnaagagctgntcncctcccncctcctntgtgtgtactactacgctcttntagacagcacgacgacacacggaggtaggagnn

      Comment

      Latest Articles

      Collapse

      • seqadmin
        Essential Discoveries and Tools in Epitranscriptomics
        by seqadmin




        The field of epigenetics has traditionally concentrated more on DNA and how changes like methylation and phosphorylation of histones impact gene expression and regulation. However, our increased understanding of RNA modifications and their importance in cellular processes has led to a rise in epitranscriptomics research. “Epitranscriptomics brings together the concepts of epigenetics and gene expression,” explained Adrien Leger, PhD, Principal Research Scientist...
        04-22-2024, 07:01 AM
      • seqadmin
        Current Approaches to Protein Sequencing
        by seqadmin


        Proteins are often described as the workhorses of the cell, and identifying their sequences is key to understanding their role in biological processes and disease. Currently, the most common technique used to determine protein sequences is mass spectrometry. While still a valuable tool, mass spectrometry faces several limitations and requires a highly experienced scientist familiar with the equipment to operate it. Additionally, other proteomic methods, like affinity assays, are constrained...
        04-04-2024, 04:25 PM

      ad_right_rmr

      Collapse

      News

      Collapse

      Topics Statistics Last Post
      Started by seqadmin, 04-25-2024, 11:49 AM
      0 responses
      20 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-24-2024, 08:47 AM
      0 responses
      20 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-11-2024, 12:08 PM
      0 responses
      62 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 10:19 PM
      0 responses
      61 views
      0 likes
      Last Post seqadmin  
      Working...
      X