Diagenode

NGSライブラリー調製

ChIP-seqライブレリー調製

ChIP後に回収できるDNA量に限りがあると、ChIP後のライブラリー調製を容易に行えません。ChIP-seqに最適なソリューションを提供するために、回収されるDNAのインプット量が低い場合でも信頼性の高いライブラリー調製を可能にする、特別に設計されたMicroPlex Library Preparation KitをDiagenodeは開発しました。 iDeal Library Preparation Kitは、DNAのインプット量が標準でも使用することもできます。また、ライブラリー調製キットとiDeal ChIP-seqおよびTrue MicroChIPキットを組み合わせてご使用頂くことで、最高のChIP-seq結果をお届けします。

DiagenodeのMicroPlex Library Preparationキットは、ChIP-seqサンプルにおいて広範囲に検証されています。 生成されたライブラリーは、シングルエンドまたはペアエンドのシーケンスと互換性があり、MicroPlexケミストリー(ステムループアダプターを使用)は、最小のインプット量から分子の複雑さの高いDNAライブラリーを生成するために特別に開発および最適化されています。 ライブラリーの調製に必要な断片化二本の鎖DNAは、わずか50 pg〜50 ngです。 3ステップのワークフロー全体の所要時間は約2時間で、単一のチューブまたはウェルで行われます。 取り扱いエラーや貴重なサンプルの損失を防ぐために、中間精製ステップやサンプル転送を省略した設計が特徴です。

特徴

  • サンプル: 断片化されたdsDNA(二本鎖DNA)
  • 低インプット量: 50 pg – 50 ng
  • 高速プロトコール: 約2時間
  • 簡単な処理: 1つのチューブに3つのステップ
  • 中間精製なし
  • シーケンス技術: イルミナ
  • 選べる手動および自動プロトコール

用途

  • ChIP由来のDNAからのChIP-seqライブラリー調製
  • 低量インプットDNAシーケンス

2種類からお選びいただけます:

The iDeal Library Preparation Kit reliably converts DNA into indexed libraries for next-generation sequencing, with input amounts down to 5 ng. Our kit offers a simple and fast workflow, high yields, and ready-to-sequence DNA on the Illumina platform.

Features

  • Sample: Fragmented dsDNA
  • Input: 5 ng – 1 µg
  • Fast protocol: 3 hours
  • Easy processing: 3 steps
  • Indexing: single indexes for multiplexing up to 24 samples
  • Manual and automated protocols available
  • Sequencing technology: Illumina

Applications

  • MeDIP-seq library prep
  • Genomic DNA sequencing
  • High input ChIP-seq
Cat. No. Product Format
C05010020 iDeal Library Preparation Kit x24 (incl. Index Primer Set 1) 24 rxns
C05010021 Index Primer Set 2 (iDeal Lib. Prep Kit x24) 24 rxns

How to choose your library preparation kit?

Application

ChIP-seq library prep
Low input DNA sequencing

MeDIP-seq library prep
Genomic DNA sequencing
High input ChIP-seq

DNA input

Low
(50 pg – 50 ng)

Standard
(5 ng – 1 µg)

Multiplexing

Up to 384 samples

Up to 48 samples

Up to 24 samples

Indexes

Dual indexes (DI)

Single indexes (SI)

Single indexes (SI)

Kit

MicroPlex Library Preparation Kit v3
(dual indexes not included in the kit)

Kit
C05010001 - 48 rxns
C05010002 - 96 rxns

Dual indexes
C05010003 - 24 DI/ 48 rxns
C05010004 - Set I 96 DI/ 96 rxns
C05010005 - Set II 96 DI/ 96 rxns
C05010006 - Set III 96 DI/ 96 rxns
C05010007 - Set IV 96 DI/ 96 rxns

MicroPlex Library Preparation Kit v2
(single indexes included in the kit)

C05010012 - 12 SI/ 12 rxns
C05010013 - 12 SI/ 48 rxns
C05010014 - 48 SI/ 48 rxns

iDeal Library Preparation Kit
(Set 1 of indexes included in the kit)

C05010020 - 12 SI/ 24 rxns

Index Primer Set 2

C05010021 - 12 SI/ 24 rxns

Combined chromatin immunoprecipitation and next-generation sequencing (ChIP-seq) has become the gold standard to investigate genome-wide epigenetic profiles. However, ChIP from a limited amount of cells has been a challenge. Here we provide a complete and robust workflow solution for successful ChIP-seq from small numbers of cells using the True MicroChIP kit and MicroPlex Library Preparation kit.

APPLICATION NOTE

ChIP efficiency on 10,000 cells

From minuscule amounts to magnificent results:
reliable ChIP-seq data from 10,000 cells with the True MicroChIP™ and the MicroPlex Library Preparation™ kits.

DOWNLOAD
APPLICATION NOTE

Quality control check of a ChIP-seq library on the Fragment Analyzer. High Efficiency ChIP performed on 10,000 cells

Best Workflow Practices for ChIP-seq Analysis with Small Samples

DOWNLOAD

MicroPlex Library Preparation Kit

  1. Can I use the available Illumina primers and validate them with the MicroPlex Kit v2?
    Although the final flanking sequences of MicroPlex are the same as those used by Illumina, the PCR primers are not identical and part of them is supplied with the buffer. For this reason Illumina primers will not work as substitute.

  2. The BioAnalyzer profile of purified library shows the presence of low molecular weight peaks (primers/adaptors) in the samples. Should I re- purify the samples or they can be used directly to the sequencing? If the second purification is recommended, which ratio sample/AMPure beads should I use?
    You can do a second round of purification using 1:1 ratio of AMPure beads to sample and this should get rid of the majority of the dimers.

  3. I am going to use the MicroPlex Library Preparation Kit v2 on ChIP samples . Our thermocycler has ramp rate 1.5°/s max while the protocol recommends using a ramp rate 3 to 5°/s. How would this affect the library prep?
    We have not used a thermocycler with a ramp rate of 1.5 °C, which seems faster than most of thermocyclers. Too fast of a ramp rate may affect the primer annealing and ligation steps.

  4. What is the function of the replication stop site in the adapter loops?
    The replication stop site in the adaptor loops function to stop the polymerase from continuing to copy the rest of the stem loop.

  5. I want to do ChIP-seq. Which ChIP-seq kit can I use for sample preparation prior to Microplex Library Preparation Kit v2?
    In our portfolio there are several ChIP-seq kits compatible with Microplex Library Preparation Kit v2. Depending on your sample type and target studied you can use the following kits: iDeal ChIP-seq Kit for Transcription Factors (Cat. No. C01010055), iDeal ChIP-seq Kit for Histones (Cat. No. C01010051), True MicroChIP kit (Cat. No. C01010130), Universal Plant ChIP-seq Kit (Cat. No. C01010152). All these kits exist in manual and automated versions.

  6. Is Microplex Library Preparation Kit v2 compatible with exome enrichment methods?
    Microplex Library Preparation Kit v2 is compatible with major exome and target enrichment products, including Agilent SureSelect®, Roche NimbleGen® SeqCap® EZ and custom panels.

  7. What is the nick that is mentioned in the kit method overview?
    The nick is simply a gap between a stem adaptor and 3’ DNA end, as shown on the schema in the kit method overview.

  8. Are the indexes of the MicroPlex library preparation kit v2 located at i5 or i7?
    The libraries generated with the MicroPlex kit v2 contain indices located at i7.

  9. Is there a need to use custom index read primers for the sequencing to read the 8nt iPCRtags?
    There is no need for using custom Sequencing primer to sequence MicroPlex libraires. MicroPlex libraries can be sequenced using standard Illumina Sequencing kits and protocols.

  10. What is the advantage of using stem-loop adapter in the MicroPlex kit?
    There are several advantages of using stem-loop adaptors. First of all, stem-loop adaptors prevent from self-ligation thus increases the ligation efficiency between the adapter and DNA fragment. Moreover, the background is reduced using ds adaptors with no single-stranded tails. Finally, adaptor-adaptor ligation is reduced using blocked 5’ ends.

IDeal Library Preparation Kit

  1. Are the index from the iDeal library Prep kit compatible with the MicroPlex library prep kit?
    No, it is important to use only the indexes provided in the MicroPlex kit to ensure proper library preparation with this kit

製品

カタログ番号製品フォーマット価格
C05010003 24 Dual indexes for MicroPlex Kit v3 /48 rxns
Diagenode’s MicroPlex Library Preparation Kits v3 have been extensively validated for ChIP-seq library prep from ChIP-derived DNA. The kit MicroPlex v3...
48 rxns $380.00
C05010004 96 Dual indexes for MicroPlex Kit v3 – Set I /96 rxns
Diagenode’s MicroPlex Library Preparation Kits v3 have been extensively validated for ChIP-seq library prep from ChIP-derived DNA. The kit MicroPlex v3...
96 rxns $1,155.00
C05010005 96 Dual indexes for MicroPlex Kit v3 – Set II /96 rxns
Diagenode’s MicroPlex Library Preparation Kits v3 have been extensively validated for ChIP-seq library prep from ChIP-derived DNA. The kit MicroPlex v3...
96 rxns $1,155.00
C05010006 96 Dual indexes for MicroPlex Kit v3 – Set III (96 rxns)
Diagenode’s MicroPlex Library Preparation Kits v3 have been extensively validated for ChIP-seq library prep from ChIP-derived DNA. The kit MicroPlex v3...
96 rxns $1,155.00
C05010007 96 Dual indexes for MicroPlex Kit v3 – Set IV /96 rxns
Diagenode’s MicroPlex Library Preparation Kits v3 have been extensively validated for ChIP-seq library prep from ChIP-derived DNA. The kit MicroPlex v3...
96 rxns $1,155.00
C05010001 MicroPlex Library Preparation Kit v3
Diagenode’s MicroPlex Library Preparation Kits v3 have been extensively validated for ChIP-seq samples and are optimized to generate DNA libraries with...
48 rxns $1,400.00
C05010020 iDeal Library Preparation Kit x24 (incl. Index Primer Set 1)
The iDeal Library Preparation Kit reliably converts DNA into indexed libraries for next-generation sequencing, with input amounts down to 1 ng. Our kit off...
24 rxns $925.00
C05010021 Index Primer Set 2 (iDeal Library Preparation Kit x24)
The Index Primer Set 2 (iDeal Library Preparation Kit x24) contains an additional set of 12  indexes to be used in combination with the iDeal Library ...
24 rxns $200.00
C05010012 MicroPlex Library Preparation Kit v2 (12 indices)
ChIP-seq専用に最適化 MicroPlex Library Preparation™kit は、ChIP-seq専用に検証されていて、ピコグラム単位の入力だけで索引付きライブラリー調製できる唯一のキットです。 True MicroChIP kitと組み合わせることで、わずか10,000細胞でC...
12 rxns $800.00
C05010014 MicroPlex Library Preparation Kit v2 (48 indices)
Specifically optimized for ChIP-seqThe MicroPlex Library Preparation™ kit is the only kit on the market which is validated for ChIP-seq and which allow...
48 rxns $2,800.00

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