Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Illumina MiSeq 16S sequencing EMP protocol questions

    Hi, I'm going to do some 16S barcoded PCR following the EMP standard protocol here (http://www.earthmicrobiome.org/proto...standards/16s/).

    The PCR potocol seems straight-forward. However, I am confused about what they say this on the website -- " For running these libraries on the MiSeq and HiSeq, please make sure you read the supplementary methods of Caporaso et al. (2012). You will need to make your sample more complex by adding 5–10% PhiX to your run."

    I have a few questions:

    1>I google the Caporaso paper and it seems PhiX is some kind of control?

    2> At which step, should I add this PhiX? Do I have to add it or it is optional? Do I need to add this into my PCR master mix? -- add it in to every samples?

    3>5-10% PhiX is the final concentration of PhiX or the volume of concentration the original PhiX?

    4> Where can I order the PhiX or Is this a kit?

  • #2
    PhiX is the control spike in from Illumina, which you can order from them. It is recommended for use with low diversity libraries and is added following the library denature and dilution. More info can be found here: http://sapac.support.illumina.com/co...5039740-10.pdf

    Comment

    Latest Articles

    Collapse

    • seqadmin
      Strategies for Sequencing Challenging Samples
      by seqadmin


      Despite advancements in sequencing platforms and related sample preparation technologies, certain sample types continue to present significant challenges that can compromise sequencing results. Pedro Echave, Senior Manager of the Global Business Segment at Revvity, explained that the success of a sequencing experiment ultimately depends on the amount and integrity of the nucleic acid template (RNA or DNA) obtained from a sample. “The better the quality of the nucleic acid isolated...
      03-22-2024, 06:39 AM
    • seqadmin
      Techniques and Challenges in Conservation Genomics
      by seqadmin



      The field of conservation genomics centers on applying genomics technologies in support of conservation efforts and the preservation of biodiversity. This article features interviews with two researchers who showcase their innovative work and highlight the current state and future of conservation genomics.

      Avian Conservation
      Matthew DeSaix, a recent doctoral graduate from Kristen Ruegg’s lab at The University of Colorado, shared that most of his research...
      03-08-2024, 10:41 AM

    ad_right_rmr

    Collapse

    News

    Collapse

    Topics Statistics Last Post
    Started by seqadmin, Today, 06:37 PM
    0 responses
    6 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, Today, 06:07 PM
    0 responses
    6 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 03-22-2024, 10:03 AM
    0 responses
    49 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 03-21-2024, 07:32 AM
    0 responses
    66 views
    0 likes
    Last Post seqadmin  
    Working...
    X