Diagenode

ChIPの成功に最も重要なステップは細胞の固定と溶解、そしてクロマチンの断片化の両方です。 研究者の中には、これらのステップの両方の重要な性質を見過すことがあります。断片化工程の不一致を解消するDiagenodeのPicoruptor®は、最先端の超音波ACT(Adaptive Cavitation Technology)を駆使し、効率的にクロマチンを断片します。ACTは、穏やかかつ非常に有効な断片力を用いて、ChIP実験に必須の高いIP効率および感度を持つ最高のクロマチン品質を可能にします。

さらに、Picoruptor®は、タンパク質-DNA複合体が高感度で偏りのない正確なChIPのために十分に保存されるよう、熱分解からクロマチン断片を保存する正確に制御された温度環境を提供します。



Diagenode Picoruptor®は、qPCRやChIP-seqなど、最適に断片化された偏りのないクロマチンが必須な多くのアプリケーションにぴったりのクロマチン断片を実現します。

Panel A, 10 µl volume: Chromatin samples are sheared for 10, 20 and 30 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.1 ml Bioruptor® Microtubes (Cat. No. B01200041). Panel B, 100 µl volume: Chromatin samples are sheared for 10 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.65 ml Bioruptor® Microtubes (Cat. No. WA-005-0500). Panel C, 300 µl volume: Chromatin samples are sheared for 5, 10 and 15 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using using 1.5 ml Bioruptor microtubes (Cat. No. C30010016). Prior to de-crosslinking, samples are treated with RNase cocktail mixture at 37°C during 1 hour. The sheared chromatin is then de-crosslinked overnight and phenol/chloroform purified as described in the kit manual. 10 µl of DNA (equivalent of 500, 000 cells) are analyzed on a 2% agarose gel (MW corresponds to the 100 bp DNA molecular weight marker).
 

Diagenodeは、完全なクロマチンを確実にするために、SDS濃度を変えて検証されたChromatin Shearing Optimizationキットを使用することを推奨しています。

SDS concentration

< 0.1%

0.2%

0.5%

1%

Nuclei isolation

Yes

Yes

Yes

No

Allows for shearing of... cells

100 million cells

100 million cells

100 million cells

100 million cells

Corresponding to shearing buffers from

iDeal ChIP-seq kit for Histones

iDeal ChIP-seq Kit for Transcription Factors

HighCell# ChIP kit

True MicroChIP kit

LowCell# ChIP kit

 

クロマチン断片サービスの推奨・関連製品

  • Bioruptor-pico-next-gen-sequencing
    B01060010
    Picoruptor ® Sonicator
  • some alt
    B01020001
    Bioruptor® Plus sonication device
  • default alt
    C01019027
    ChIP Cross-link Gold
  • default alt
    C01020014
    Chromatin Shearing Optimization Kit (Universal ...
  • ChIP kit icon
    C01020010
    Chromatin shearing optimization kit - Low SDS (...
  • ChIP kit icon
    C01020013
    Chromatin shearing optimization kit – Low SDS (...
  • Bioruptor One
    B01070001
    Diagenode One sonication device
  • some alt
    C30010015
    0.1 ml Picoruptor Microtubes
  • some alt
    B01200041
    0.1 ml tube holder & tube adaptors for Picoruptor
  • some alt
    C30010013
    0.5 ml Bioruptor® Plus Microtubes
  • Product image
    B01200043
    0.5/0.65 ml tube holder for Picoruptor
  • some alt
    C30010011
    0.65 ml Picoruptor Microtubes
  • some alt
    C30010010-300
    1.5 ml Bioruptor® Plus TPX microtubes
  • some alt
    C30010010-50
    1.5 ml Bioruptor® Plus TPX microtubes
  • some alt
    C30010016
    1.5 ml Picoruptor Microtubes with Caps
  • some alt
    B01200040
    1.5 ml tube holder for Picoruptor
  • some alt
    B01200011
    1.5 ml tube holder for Bioruptor® Standard ...
  • some alt
    B01200012
    10 ml tube holder for Bioruptor® Standard & Plus
  • some alt
    C30010009
    15 ml Bioruptor® Plus TPX tubes
  • some alt
    C01020031
    15 ml Picoruptor Tubes & sonication beads
  • some alt
    B01200016
    15 ml sonication accessories for Picoruptor
  • some alt
    B01200013
    15 ml tube holder for Bioruptor® Standard &...
  • ChIP kit icon
    C01020012
    Chromatin Shearing Optimization Kit - High SDS ...
  • ChIP kit icon
    C01020011
    Chromatin Shearing Optimization Kit - Medium SD...
  • ChIP kit icon
    C01020022
    Shearing Optimization kit

関連するプロトコル

Chromatin shearing - Guide for successful chromatin preparation using the Bioruptor® Pico
Chromatin immunoprecipitation (ChIP) is a common tool used to study protein-DNA interactions in c...
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Chromatin shearing from tissue protocol using Diagenode’s Chromatin shearing optimization kit - Low SDS and Bioruptor® Standard, Plus or Pico
This protocol describes the chromatin preparation from fresh or frozen tissues. The isolated chro...
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ChIP - The Ultimate Guide for Chromatin Shearing Optimization with Bioruptor® Standard and Plus
A review of the critical points for chromatin shearing.
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