Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Optimal RNA prep from whole mouse tissue

    I have an experiment planned in which RNA samples will be collected from total mouse tissue (eg whole kidney, whole brain, etc).
    I plan to use Qiagen RNeasy kit for RNA prep and then do downstream library prep and RNASeq.

    Since I want to get data on whole tissue RNA content (and I do realize I will only see very large-scale changes since any change occurring in a subset of cells will likely be invisible to me here) I would like to homogenize the entire tissue (and not just a portion). I will need to collect the entire tissue at the end of the in vivo part of the experiment and later process for RNA

    What would be your recommendation on tissue preservation (given the size of a whole mouse organ)? What would be the best way to perform initial homogenization of the entire tissue? After this I can subsample a portion of the total homogenized sample for downstream RNA preparation and sequencing

    Thanks
    Noa

  • #2
    I would take 5mm x 5mm blocks of tissue and snap freeze them.
    Macrodissect and homogenise in RLT using a hand homogeniser, or use a bead mill homogeniser.
    Last edited by Marc_Jones; 03-14-2019, 01:45 AM. Reason: Clarity

    Comment

    Latest Articles

    Collapse

    • seqadmin
      Best Practices for Single-Cell Sequencing Analysis
      by seqadmin



      While isolating and preparing single cells for sequencing was historically the bottleneck, recent technological advancements have shifted the challenge to data analysis. This highlights the rapidly evolving nature of single-cell sequencing. The inherent complexity of single-cell analysis has intensified with the surge in data volume and the incorporation of diverse and more complex datasets. This article explores the challenges in analysis, examines common pitfalls, offers...
      06-06-2024, 07:15 AM
    • seqadmin
      Latest Developments in Precision Medicine
      by seqadmin



      Technological advances have led to drastic improvements in the field of precision medicine, enabling more personalized approaches to treatment. This article explores four leading groups that are overcoming many of the challenges of genomic profiling and precision medicine through their innovative platforms and technologies.

      Somatic Genomics
      “We have such a tremendous amount of genetic diversity that exists within each of us, and not just between us as individuals,”...
      05-24-2024, 01:16 PM

    ad_right_rmr

    Collapse

    News

    Collapse

    Topics Statistics Last Post
    Started by seqadmin, Yesterday, 06:58 AM
    0 responses
    13 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 06-06-2024, 08:18 AM
    0 responses
    20 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 06-06-2024, 08:04 AM
    0 responses
    18 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 06-03-2024, 06:55 AM
    0 responses
    13 views
    0 likes
    Last Post seqadmin  
    Working...
    X