Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • Selbbub
    Junior Member
    • Mar 2012
    • 1

    Sonication woes

    Hi everyone,

    I am having some difficulties optimising sonication and would like some advice I am trying to sonicate approx. 12 million cells in 1.5ml lysis buffer using a Sonics Vibra Cell. I'm currently using 40% amplitude and 10 sec bursts, however after 40x10 seconds my chromatin is an even smear from 200-2000 bp with no concentrated region. This size range so far seems to be working for regular ChIP, but obviously isn't anywhere near small enough for ChIPSeq.

    It seems like I am already sonicating a lot compared to what most other people need to do.. any suggestions?
  • jo_4
    Junior Member
    • Sep 2012
    • 1

    #2
    Hi selbbub,
    What kind of cells are you using? Does your lysis buffer have 1% SDS? Trying to reduce the volume to 600ul might work..

    Comment

    Latest Articles

    Collapse

    • GATTACAT
      Reply to Nine Things a Sample Prep Scientist Thinks About Before Sequencing
      by GATTACAT
      Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.
      07-01-2026, 11:43 AM
    • SEQadmin2
      Nine Things a Sample Prep Scientist Thinks About Before Sequencing
      by SEQadmin2


      I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.

      Here are nine questions we think about, in roughly the order they matter, before...
      06-18-2026, 07:11 AM

    ad_right_rmr

    Collapse

    News

    Collapse

    Topics Statistics Last Post
    Started by SEQadmin2, 07-02-2026, 11:08 AM
    0 responses
    26 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 06-30-2026, 05:37 AM
    0 responses
    24 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 06-26-2026, 11:10 AM
    0 responses
    24 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 06-17-2026, 06:09 AM
    0 responses
    55 views
    0 reactions
    Last Post SEQadmin2  
    Working...