Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • alisrpp
    Member
    • Dec 2010
    • 40

    Annealing index and primers for multiplexing??

    Hi,
    We are planning to do a multiplexed cDNA library to run on Illumina . We bought the indexes and primers from IDT (sequences from Illumina) but we have some questions. We were reading that Illumina changed the protocol and now it uses the index+PCRprimer (all together) instead of using the index as a "third" primer.
    We were thinking about annealing the index+primer and use it as a single one but we are not sure if it is gonna work. Does anybody have tried that?
    If so, which temperature?
    Otherwise we will use as separated things. Has anybody got good results with this?

    Thanks
  • RNAseqer
    Member
    • Sep 2010
    • 22

    #2
    ahhh...we used to do this but not anymore. Just couldn't get good annealing, we'd see multiple peaks. I recommend using the TruSeq adapters from Illumina or the DNA Adapters from Bioo which are sold separately from their library prep kit.

    Comment

    Latest Articles

    Collapse

    • SEQadmin2
      Nine Things a Sample Prep Scientist Thinks About Before Sequencing
      by SEQadmin2


      I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.

      Here are nine questions we think about, in roughly the order they matter, before...
      06-18-2026, 07:11 AM
    • SEQadmin2
      From Collection to Sequencing: Why Sample Preparation and Preservation Define Sequencing Data
      by SEQadmin2


      Data variability is still an issue in sequencing technologies despite the advances in reproducibility and accuracy of these platforms. But the problem does not originate in the sequencing itself, but in the previous steps, before the sample reaches the sequencer.


      The first step is collection, followed by preservation and sample preparation for analysis. Most scientists overlook those steps, but not being careful might just be skewing the experiment’s results.
      ...
      06-02-2026, 10:05 AM

    ad_right_rmr

    Collapse

    News

    Collapse

    Topics Statistics Last Post
    Started by SEQadmin2, 06-17-2026, 06:09 AM
    0 responses
    36 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 06-09-2026, 11:58 AM
    0 responses
    100 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 06-05-2026, 10:09 AM
    0 responses
    121 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 06-04-2026, 08:59 AM
    0 responses
    113 views
    0 reactions
    Last Post SEQadmin2  
    Working...