Header Leaderboard Ad

Collapse

What to even do with pooled 16S and ITS amplicon sequences with no barcodes?

Collapse

Announcement

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

  • What to even do with pooled 16S and ITS amplicon sequences with no barcodes?

    Hey there
    I've been working exclusively with bioinformatics for a few years and following DNA extraction, I leave all my sequencing needs to be solved by sequencing facilities. Most of my projects require both 16S and ITS sequencing and in most cases, I've received the data in separate runs. Recently, I decided to evaluate a recent company from my country and they sent me the data for both amplicons in the same file. I thought that maybe the samples were multiplexed, but to my surprise, they said they just pool the libraries and run them without indexing per target.
    Is there even a correct way to deal with this? I use a DADA2-based workflow, I thought about maybe doing a preliminary classification of each read (using just blast), then separating the targets and following my usual workflow, but this feels like a workaround I should not be taking.

  • #2
    Hi,
    you might use the primer sequences (which might be obtained from the company) to split the data in an ITS and a 16S part which can be analysed separately. Run e.g. cutadapt on the data set with the ITS primer and save the untrimmed reads in a file, which you use as an input for the trimming with the 16S primers.

    Comment

    Latest Articles

    Collapse

    • seqadmin
      Improved Targeted Sequencing: A Comprehensive Guide to Amplicon Sequencing
      by seqadmin



      Amplicon sequencing is a targeted approach that allows researchers to investigate specific regions of the genome. This technique is routinely used in applications such as variant identification, clinical research, and infectious disease surveillance. The amplicon sequencing process begins by designing primers that flank the regions of interest. The DNA sequences are then amplified through PCR (typically multiplex PCR) to produce amplicons complementary to the targets. RNA targets...
      03-21-2023, 01:49 PM
    • seqadmin
      Targeted Sequencing: Choosing Between Hybridization Capture and Amplicon Sequencing
      by seqadmin




      Targeted sequencing is an effective way to sequence and analyze specific genomic regions of interest. This method enables researchers to focus their efforts on their desired targets, as opposed to other methods like whole genome sequencing that involve the sequencing of total DNA. Utilizing targeted sequencing is an attractive option for many researchers because it is often faster, more cost-effective, and only generates applicable data. While there are many approaches...
      03-10-2023, 05:31 AM

    ad_right_rmr

    Collapse

    News

    Collapse

    Topics Statistics Last Post
    Started by seqadmin, Yesterday, 11:44 AM
    0 responses
    8 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 03-24-2023, 02:45 PM
    0 responses
    18 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 03-22-2023, 12:26 PM
    0 responses
    18 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 03-17-2023, 12:32 PM
    0 responses
    19 views
    0 likes
    Last Post seqadmin  
    Working...
    X