Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • PCR product quantification

    We plan to use uniq adaptor tagged primers for a PCR and then pool PCR products of about 5 samples together for one TruSeq index and follow the TruSeq protocol skipping fragmentation.
    Pooling similar or near equal amounts of PCR products (purified) would be a key issue. I would be grateful for reccomendations on quantification methods. And the actual risk of using nano drop for quantitication. Would I be digging my own grave if I use nano drop ? Using any other method would be an additional expense associated

  • #2
    Whether or not nanodrop is sufficient will depend on how perfect you need the pooling. If 20% varaince is fine, then nanodrop should be okay.

    If you need perfectly equal pooling, I would suggest at least Bioanalyzer/Tape Station and preferrably qPCR.

    The extra cost of those should outweigh the need to run the sample again to get sufficient reads.

    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...
      04-22-2024, 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, Today, 08:47 AM
    0 responses
    11 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-11-2024, 12:08 PM
    0 responses
    60 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-10-2024, 10:19 PM
    0 responses
    59 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-10-2024, 09:21 AM
    0 responses
    54 views
    0 likes
    Last Post seqadmin  
    Working...
    X