Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Barcoded files, a lot of variance in supposed primers, help me understand

    So, I have this file with 1,314 sequences (file1) and another file with 566 sequences (file2). Each sequence has a proper barcode. What I can't understand is that in file1 the letters following the barcode are (number is count):

    AAAGGGGCGCAGCAGGCGCG 1
    AACGGGGCGACAGCAGGCGC 1
    ACCGGGCGCAGCAGGCGCGA 1
    ACGGGCGCAGCAGGCGCGAA 8
    ACGGGCGCTAGCAGGCGCGA 1
    ACGGGGACGCAGCAGGCGGA 1
    ACGGGGCACAGCAGGCGCGA 1
    ACGGGGCACAGTAGGCGCGA 1
    ACGGGGCCAGCAGGCGCGAA 2
    ACGGGGCCGCAGCAGGCGCG 2
    ACGGGGCGAAAGCAGGCGCG 1
    ACGGGGCGAGCAGGCGCGAA 1
    ACGGGGCGCAAGCAGGCGCG 1
    ACGGGGCGCACGCAGGCGCG 1
    ACGGGGCGCAGCAAAGCGCG 1
    ACGGGGCGCAGCAAGCGCGA 3
    ACGGGGCGCAGCAAGGCGCG 3
    ACGGGGCGCAGCACGCGCGA 1
    ACGGGGCGCAGCACGGCGCC 1
    ACGGGGCGCAGCACGGCGCG 3
    ACGGGGCGCAGCACGGCGGC 4
    ACGGGGCGCAGCAGACGCGA 4
    ACGGGGCGCAGCAGGAGCGA 1
    ACGGGGCGCAGCAGGCACGA 3
    ACGGGGCGCAGCAGGCGACG 5
    ACGGGGCGCAGCAGGCGCCG 1
    ACGGGGCGCAGCAGGCGCGA 1219
    ACGGGGCGCAGCAGGCGGCG 2
    ACGGGGCGCAGCAGGCTCGA 1
    ACGGGGCGCAGCATGCGCGA 1
    ACGGGGCGCAGCTACGGCGC 3
    ACGGGGCGCAGGCAGGCGCG 1
    ACGGGGCGCAGTAGGCGCGA 1
    ACGGGGCGCAGTCACGGCGC 1
    ACGGGGCGCAGTCAGGCCGC 1
    ACGGGGCGCAGTCAGGCGCG 5
    ACGGGGCGCCGCAGGCGCGA 3
    ACGGGGCGCGCAGGCGCGAA 1
    ACGGGGCGCTAGTCAGGCGC 2
    ACGGGGCGGCAGCAGGCGCG 3
    ACGGGGGCGCAGCAGGCGCG 13
    ACGGGGGGCGCAGCAGGCGC 1
    ACGGGTCGCAGCAGCGCGAA 1
    AGTGGGGCGCAGCAGGCGCG 1
    ATGGGGCGCAGCAGGCGCGA 1
    GACGGGGCGCAGCAGGCGAC 3
    GACGGGGCGCAGCAGGCGCG 2

    However, in file2:
    AACGGGGCGTCAGCAGGGCG 1
    ACGGGCCGCAGCAGGGCGCG 1
    ACGGGCGCAGCAGGCGCGAA 2
    ACGGGCGCAGCAGGGCGCGA 151
    ACGGGCGCTAGCACGGCGGC 1
    ACGGGCGCTAGCACGGGCGG 1
    ACGGGCGCTAGCAGGCGCGA 1
    ACGGGCGCTAGCAGGCGCGG 2
    ACGGGCGCTAGCAGGGCGCG 1
    ACGGGCGCTAGCAGGGCGGC 1
    ACGGGCGGCAGCAGGGCGCG 1
    ACGGGCGTCAGCAGGACGCG 1
    ACGGGCGTCAGCAGGCGGCG 3
    ACGGGCGTCAGCAGGGCGCG 3
    ACGGGCGTCAGCAGGGCGGC 1
    ACGGGCGTCAGCAGGGGCGC 1
    ACGGGGCCAGCAGGGCGCGA 3
    ACGGGGCCGCAGCAGGGCGC 2
    ACGGGGCGAAGCAGGGCGCG 1
    ACGGGGCGAGCAGGGCGCGA 1
    ACGGGGCGCAGCAAGCGCGA 1
    ACGGGGCGCAGCAGCGCGAA 1
    ACGGGGCGCAGCAGGGCGAC 2
    ACGGGGCGCAGCAGGGCGCG 340
    ACGGGGCGCAGCAGGGCGGC 8
    ACGGGGCGCAGCAGGGGCGC 1
    ACGGGGCGCAGCAGGGTGCG 4
    ACGGGGCGCAGCAGTCGCGA 1
    ACGGGGCGCTAGCAGGGCGC 1
    ACGGGGCGCTAGCAGGGCGG 1
    ACGGGGCGTCAGCAGGGCGC 4
    ACGGGGCGTCAGCAGGGCGG 4
    ACGGGGCGTCAGCAGGGGCG 5
    ACGGGGCGTCAGCTAGGGGC 1
    ACGGGGCTAGCAGGCGCGGA 1
    ACGGGGTCGCAGCAGGGCGC 5
    ACGGGGTCGCAGCAGGGGCG 3
    ACGGGTCGCAGCAGGGCGCG 1
    ACGGGTGCAGCAGGGCGCGA 1
    ACGGGTGTCAGCAGGGCGCG 1
    ATGGGGGCGCAGCAGGGCGC 1
    CCGGGGCGCAGCAGGGCGCG 1
    GACGGGCGCAGCAGGGCGCG 1
    GACGGGGCGCAGCAGGGCGA 1
    GACGGGGCGCAGCAGGGCGC 1

    The same forward primer 'ACGGGGCGCAGCAGGCGCGA' (the most abundant seq in file1) was used for both files. It's not even present in file2. How can this be? It's as if file2 is barcoded but the primer has been removed or something?
    Last edited by rhinoceros; 10-17-2013, 05:22 AM.
    savetherhino.org

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, 04-11-2024, 12:08 PM
0 responses
59 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-10-2024, 10:19 PM
0 responses
57 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-10-2024, 09:21 AM
0 responses
53 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-04-2024, 09:00 AM
0 responses
56 views
0 likes
Last Post seqadmin  
Working...
X