Investigator IDplex Plus Kit

13 CODISコア遺伝子座、加えてD2S1338、D19S433およびアメロゲニンもスワブやペーパーから直接増幅するため

S_1084_5_GEN_V2

✓ オンライン注文による24時間年中無休の自動処理システム

✓ 知識豊富で専門的な製品&テクニカルサポート

✓ 迅速で信頼性の高い(再)注文

Investigator IDplex Plus Kit (100)

Cat. No. / ID:   381625

プライマーミックス、Fast Reaction Mix(HotStarTaq Plus DNAポリメラーゼを含む)、コントロールDNA、アレリックラダーIDplex Plus、DNA標準サイズ550(BTO)、ヌクレアーゼフリー水
KitAccessories
Investigator IDplex Plus
Matrix Standard
Reactions
100
400
Investigator IDplexキットは、法医学、ヒト個人識別、および父子鑑定における分子生物学アプリケーションでの使用を想定しています。病気の診断、予防、治療を目的としてはいません。これらの製品は、一部の国々ではご利用いただけません。

✓ オンライン注文による24時間年中無休の自動処理システム

✓ 知識豊富で専門的な製品&テクニカルサポート

✓ 迅速で信頼性の高い(再)注文

特徴

  • CODISコアを含め、16個の遺伝子座の同時増幅
  • 法医学ケースワークまたは標準サンプルに最適
  • すぐに使えるFast Reaction Mixでの簡単なプロトコール
  • マーカーの重複を低減する最適化されたプライマー設計

製品詳細

Investigator IDplex Plus Kitは、13 CODIS(Combined DNA Index System)コア遺伝子座、D2S1338、D19S433、およびアメロゲニンを網羅するヒト個人識別用のマルチプレックスSTRアッセイです。

キットは、高速サイクリング技術を使用し、およそ90分で増幅を可能にします。Investigator IDplex Plus Kitは、高感度で、微量DNAからの迅速で信頼性の高い結果が得られるため、すべての法医学アプリケーションと父子鑑定検査に適しています。ジェネティックアナライザは、BT5 Matrixで較正します。

Investigator IDplex Kitは、ISO 18385要件を満たします。

パフォーマンス

Investigator IDplex Kitは、抽出されたDNAからおよそ90分でSTRアッセイで信頼性の高い結果を出すため、高速サイクリング技術を使用します。

Fast Reaction Mixは、阻害剤耐性を向上させるために最適化された実証済みのQIAGENマルチプレックスPCRテクノロジーに基づいています。6種類の阻害剤(フミン酸、ヘマチン、タンニン酸、インジゴカルミン、カルシウム、コラーゲン)の濃度を増加させて検査したアッセイの性能は、高感度と優れた阻害剤耐性を示しています。これにより、ケースワークサンプルから信頼性の高い、高品質の結果を得ることが可能です。

Investigator IDplex Kitは、GeneAmp 9700サーマルサイクラーとApplied Biosystems 3500 Genetic Analyzerを使用して検証・評価済みです。

Investigator IDplex Plus Kitと共に使用するための標準条件下でのDNAの推奨量は、0.5 ngです。内部検証では、0.2–2 ng DNAで頑健でバランスのとれた結果、そして0.1 ng未満のDNAで信頼性の高い結果が示されました。

原理

Investigator IDplex Kitは、13個のCODIS(Combined DNA Index System)コア遺伝子座、D2S1338、D19S433、およびアメロゲニンの迅速な同時増幅のために、QIAGENの高速サイクリングPCRテクノロジーを使用しています。注意深く設計されたプライマーは、遺伝子座アレルの重複を低減し、これにより誤った解釈の可能性を最小限に抑えます。

操作手順

15個の多型常染色体STRマーカー(D2S1338、D3S1358、D5S818、D7S820、D8S1179、D13S317、D16S539、D18S51、D19S433、D21S11、CSF1PO、FGA、TH01、TPOX、およびvWA)、および性差アメロゲニンを、シングルPCRで同時増幅し、表「Investigator IDplex Plus Kit色素と対応するマーカー」にリストされているいずれかの色素で標識します。

 

Investigator IDplex Plus Kit色素と対応するマーカー
色素 マーカー
6-FAM アメロゲニン TH01 D3S1358 vWA D21S11
BTG TPOX D7S820 D19S433 D5S818 D2S1338
BTY D16S539 CSF1PO D13S317 FGA  
BTR D18S51 D8S1179      

増幅した遺伝子座は、それからキャピラリー電気泳動します。結果は、GeneMapper ID-Xなどの専用ソフトウェアを使用して分析できます。詳細は、表「技術仕様」を参照してください。

アプリケーション

Investigator IDplex Plusキットは、血液、口腔スワブ、法医学的染み、または標準サンプルからDNAプロファイルを迅速に、かつ高い信頼性で作成するためのものであり、以下に最適です。

  • 犯罪データベース、法医学的標準またはケースワークサンプル
  • 父子鑑定検査
  • 集団遺伝学的調査
  • その他のヒト個人識別検査

 

技術仕様
機能 Investigator IDplex Plus Kit
DNAインプット量 最大15 µl
反応当りの容量 25 µl
マトリックス BT5
蛍光色素 6-FAM、BTG、BTY、BTO
ジェネティックアナライザ ABI PRISM 3100/3100 Avant、Applied
Biosystems 3130/3130xl、またはApplied
Biosystems 3500 Genetic Analyzers
分析用ソフトウェア Applied Biosystems GeneMapper ID、または
GeneMapper ID-X

リソース

パンフレット (3)
Advanced sample collection, automation and STR solutions for kinship testing

Streamline your workflow with QIAGEN solutions for human identity and forensic testing

キットハンドブック (1)
クイックスタートプロトコール (1)
テンプレートファイル (3)
For further information on which version of the v11 template files is suitable for your needs, please refer to the Template File Info Note.
For further information on which version of the v11 template files is suitable for your needs, please refer to the Template File Info Note.
テクニカルインフォメーション (3)

The most comprehensive and versatile coverage of current international STR standards for capillary electrophoresis analysis



Quality initiatives for human identity testing and forensics 
Technical Information and Important Notes (1)
MSDS (2)
Download Safety Data Sheets for QIAGEN product components.
Download Safety Data Sheets for QIAGEN product components.
Safety Data Sheets (1)
Certificates of Analysis (1)

FAQ

Is the Fast Reaction Mix of the Investigator IDplex Plus Kit ready to use?
Yes. The Fast Reaction Mix of the Investigator IDplex Plus Kit contains HotStarTaq Plus enzyme, Mg2+, and dNTPs in a pre-optimized concentration. Only Investigator IDplex Plus Primer Mix and template DNA need to be added. Nuclease-free Water should be added to complete the reaction volume.
FAQ ID -3008
What is the safest procedure to add FTA punches to the PCR reactions of the Investigator IDplex GO! Kit?

We recommend first adding master mix to the PCR plate and then adding the punch to avoid punches getting lost or mixed up due to electrostatic effects. Furthermore, we strongly recommend spinning down the PCR plate after setup to ensure all punches get fully submerged.

FAQ ID -3096
Can the Investigator IDplex GO! PCR protocol be run on different PCR endpoint cyclers?
Yes. The protocol is generic and has successfully been tested on various commonly used instruments.
FAQ ID -3089
Can the PCR protocol of the Investigator IDplex Plus Kit be run on different PCR endpoint cyclers?
Yes. The protocol of the Investigator IDplex Plus Kit is generic and has been successfully tested on various commonly used instruments.
FAQ ID -3009
What is the maximum DNA input volume that can be used in the Investigator IDplex Plus Kit?
15 µl input volume can be used with the Investigator IDplex Plus Kit.
FAQ ID -3014
Can additional positive controls be used for the Investigator IDplex GO! Kit?

Amplification of Control DNA 9948 is appropriate as a PCR performance check. However, if you have access to reference samples at constant quality (for example, swabs or FTA donated by lab personnel), these samples may be used as additional in process controls.

FAQ ID -3094
Which filter set should be used with Investigator IDplex Plus Kit?
If you are using an ABI PRISM 3100-Avant/3100 or 3130/3130xl Genetic Analyzer, please use Any5Dye, if available. If you are using the Applied Biosystems 3500/3500xL Genetic Analyzer, please use filter set AnyDye. Otherwise, filter set G5 should be used. If you are using an ABI PRISM 310 Genetic Analyzer, filter set G5 should be used.
FAQ ID -3017
Does the Investigator IDplex Plus assay contain an internal control?
No. Unlike some other QIAGEN STR products (e.g., the Investigator Argus Y-12 QS Kit) there is no QS quality sensor included in the Investigator IDplex Plus Kit.
FAQ ID -3011
Is there a need to compensate for additional liquid volume added with the positive control DNA, or a buccal swab lysate in the Investigator IDplex GO! Kit?
No. Up to 2 µl can be added on top of a 25 µl reaction.
FAQ ID -3088
Is it possible to increase the amount of sample to obtain higher signals using the Investigator IDplex GO! Kit?
We do not recommend using more than one punch of 1.2 mm diameter for blood or buccal cells on FTA, or 2 µl of a crude buccal swab lysate. To adapt protocols to the type and quality of samples used in your laboratory, and to the CE instrumentation in use, adjust PCR cycle numbers and CE injection conditions.
FAQ ID -3090
Do you recommend using indicator cards for buccal cells on FTA with Investigator IDplex GO! Kit?
Yes. Use of indicator cards helps to ensure sample is taken from an appropriate area of the FTA card. This may significantly improve success rates compared to procedures using non-indicator cards.

 

FAQ ID -3095
What is the optimal sample DNA concentration for the Investigator IDplex Plus Kit?
The optimum sample DNA input range of the Investigator IDplex Plus Kit is between 200–500 pg. However, reliable results can be obtained with less than 100 pg. For higher amounts of input DNA, consider adjusting the PCR cycle number.
FAQ ID -3015
Can the allelic ladder of the Investigator IDplex Plus Kit be used for analysis of Investigator IDplex GO! Kit?

No. Amplification products of marker FGA have a slightly different size between IDplex Plus and IDplex GO! kits due to a change in the fluorescent dye linker used. Use of the IDplex Plus ladder will frequently cause off ladder calls for FGA alleles.

 

FAQ ID -3093
Is the Fast Reaction Mix ready to use in the Investigator IDplex GO?

Yes. The Fast Reaction Mix contains Taq enzyme, Mg2+, and dNTPs in a pre-optimized concentration. Only IDplex GO! Primer Mix and sample needs to be added.

FAQ ID -3087
Do I need to evaluate optimal PCR cycle numbers for the Investigator IDplex GO! Kit with my samples?

Yes. Sample materials used for direct amplification vary with regard to quality and quantity of DNA present, depending on sampling materials, procedures, and storage conditions. In order to obtain the highest possible success rates in routine use, it is essential to run a representative batch of samples for evaluation of optimal conditions. Furthermore, we recommend revisiting this aspect after a certain period of routine use to ensure the evaluated conditions are optimal and re-adjust if necessary.

FAQ ID -3091
Have primer sequences of the Investigator IDplex Plus Kit been changed compared to the former Investigator IDplex assay?
No, the primer site locations of the markers in the Investigator IDplex Plus Kit have not been changed. Only minor changes have been introduced for some markers, e.g., to improve A addition to the PCR products.
FAQ ID -3012
What is the correct matrix to use with Investigator IDplex Plus Kit?
Matrix BT5 should be used with Investigator IDplex Plus Kit.
FAQ ID -3016
What fluorescent labels are 6-FAM, BTG, BTY, BTR, and BTO in Matrix Standard BT5 single or multi.cap?

The BT5 matrix standard is designed for use with our 5-dye Investigator STR Assays. We do not disclose the containing dyes.

FAQ ID - 3364
Is it possible to extend the number of PCR cycles to increase sensitivity of the Investigator IDplex Plus Kit?
Yes. For example, cycle numbers can be increased to 32 or 34 in order to obtain higher signals for very diluted samples. However, please note that this promotes the amplification of nonspecific products or amplification from spurious background DNA.
FAQ ID -3010
What does the Investigator IDplex Plus Kit contain?
The Investigator IDplex Plus Kit contains Primer mix, Fast Reaction Mix, Control DNA allelic ladder IDplex Plus, DNA size standard 550 (BTO), and nuclease-free water.
FAQ ID -3013
Where can I find the template files for the Investigator IDplex Plus Kit?
The template files for the Investigator IDplex Plus Kit for various softwares and analyzers can be downloaded from the Resource tab on the product webpage.
FAQ ID -3019
Have primer sequences in the Investigator IDplex GO! Kit been changed compared to the Investigator IDplex Plus assay?

No, primer site locations of markers have not been changed. Two additional SNP primers have been added to allow balanced amplification of vWA and D16S539 alleles carrying known primer binding site mutations. The mutations covered were observed in concordance studies in samples of African origin at an elevated level. The labelled FGA forward primer has been slightly changed with regard to the linker of the fluorescent dye. The change does not affect PCR amplification, but causes a slight size shift during capillary electrophoresis. The allelic ladder therefore has been changed accordingly.

FAQ ID -3092
Which settings for the filter set Any5Dye must be adjusted before running the spectral calibration for Investigator IDplex Plus Kit?

For Any5Dye filter set with the Investigator IDplex Plus Kit, please use:

  • Condition Bound: Lower 1.0 – Upper 20
  • Sensitivity: 0.1
  • Quality Value: 0.80
FAQ ID -3018