Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Nextera XT vs TruSeq PCR free

    I have previously used the TruSeq DNA kit (only once) and now need to move on to use either Nextera XT or TruSeq PCR free kits and perform the whole library prep alone for the first time.
    I would greatly welcome opinions on which of the two kits have given better results for small genomes (bacterial)

    2)
    For Nextera XT, is it better not to use the bread normalization and do the traditional quantifications? There seem to be mixed experiences for using bead normalizations.

    3) is there anyone using Qubit alone for library quantification? Or is it better to use KAPA qPCR instead or both?

    4) This is probably a very naive question,
    for Nextera XT with bacterial genomes, is it necessary to sonicate/ nebulize to get a larger size fragment and then use the kit based tagmentation than the kit alone.

    Thanks a lot

  • #2
    1) For bacterial genome it would not make much difference in result. The consideration would be cost and availability of DNA input amount.

    2) qPCR based quantification (not using normaliser beads) will increase chance of even number of read output for samples.

    3) KAPA qPCR certainly is better than Qubit. Qubit only measures mass of DNA not cluster ready portion of library.

    4) This question is a bit unclear to me. Using high molecular weight input DNA (without sonication) will give more consistent results. Sonication is not in the protocol and I do not see any reason for sonication.
    Last edited by nucacidhunter; 06-09-2014, 01:30 AM.

    Comment

    Latest Articles

    Collapse

    • seqadmin
      Understanding Genetic Influence on Infectious Disease
      by seqadmin




      During the COVID-19 pandemic, scientists observed that while some individuals experienced severe illness when infected with SARS-CoV-2, others were barely affected. These disparities left researchers and clinicians wondering what causes the wide variations in response to viral infections and what role genetics plays.

      Jean-Laurent Casanova, M.D., Ph.D., Professor at Rockefeller University, is a leading expert in this crossover between genetics and infectious...
      09-09-2024, 10:59 AM
    • seqadmin
      Addressing Off-Target Effects in CRISPR Technologies
      by seqadmin






      The first FDA-approved CRISPR-based therapy marked the transition of therapeutic gene editing from a dream to reality1. CRISPR technologies have streamlined gene editing, and CRISPR screens have become an important approach for identifying genes involved in disease processes2. This technique introduces targeted mutations across numerous genes, enabling large-scale identification of gene functions, interactions, and pathways3. Identifying the full range...
      08-27-2024, 04:44 AM

    ad_right_rmr

    Collapse

    News

    Collapse

    Topics Statistics Last Post
    Started by seqadmin, Today, 06:25 AM
    0 responses
    13 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, Yesterday, 01:02 PM
    0 responses
    12 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 09-18-2024, 06:39 AM
    0 responses
    14 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 09-11-2024, 02:44 PM
    0 responses
    14 views
    0 likes
    Last Post seqadmin  
    Working...
    X