Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Nextera XT library preparation

    I am using Nextera XT for library preparation.
    The kit suggests 12 cycles of PCR after tagmentation.
    If I run 5 ul of PCR product after 12 cycles I do not see any smear on my 1.5% agarose gel. If I run 13 cycles then I see the smear.
    I am wondering if this is normal or if I should see a smear after 12 cycles.
    I will try to go to the Bioanalyzer to see my library but I am not sure it will give me a better answer.
    Any help or comment could be really useful.

  • #2
    I do not run my libraries on gel to check, so I do not know if one should see smear or not. If you see smear after 13 cycles by running 5 ul of PCR, then using 10 ul after 12 cycles should give similar results.

    Comment


    • #3
      This is normal. Many people modify the protocol to increase the number of PCR cycles like this.

      Comment

      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 on Modified Bases...
        Today, 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
      37 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 10:19 PM
      0 responses
      41 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 09:21 AM
      0 responses
      35 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-04-2024, 09:00 AM
      0 responses
      54 views
      0 likes
      Last Post seqadmin  
      Working...
      X