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RT2 SYBR® Green FAST Mastermixesは分子生物学的アプリケーション用であり、疾病の診断、予防、あるいは治療に使用することはできません。
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RT² SYBR Green ROX FAST Mastermix (2)

Cat. No. / ID:  330620

Contains 2 x 1.35 ml tubes: for 12 x 100-well ring discs on the QIAGEN Rotor-Gene Q or 250 x 20 µl reactions
¥37,000
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RT² SYBR Green FAST Mastermix (8)

Cat. No. / ID:  330601

Contains 8 x 1.35 ml tubes: for 8 x 100-well ring discs on the QIAGEN Rotor-Gene Q or 1000 x 20 µl reactions
¥136,000
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RT² SYBR Green ROX FAST Mastermix (24)

Cat. No. / ID:  330623

Contains 24 x 1.35 ml tubes: for 24 x 100-well ring discs on the QIAGEN Rotor-Gene Q or 3000 x 20 µl reactions
¥306,000
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RT² SYBR Green ROX FAST Mastermix (25 ml)

Cat. No. / ID:  330629

Contains 1 bottle of 25 ml mastermix: for 24 x 100-well ring discs on the QIAGEN Rotor-Gene Q or 2500 x 20 µl reactions
¥271,000
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RT² SYBR Green FAST Mastermix (12)

Cat. No. / ID:  330602

Contains 12 x 1.35 ml tubes: for 12 x 100-well ring discs on the QIAGEN Rotor-Gene Q or 1500 x 20 µl reactions
¥199,000
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RT² SYBR Green ROX FAST Mastermix (8)

Cat. No. / ID:  330621

Contains 8 x 1.35 ml tubes: for 8 x 100-well ring discs on the QIAGEN Rotor-Gene Q or 1000 x 20 µl reactions
¥136,000
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RT² SYBR Green FAST Mastermix (2)

Cat. No. / ID:  330600

Contains 2 x 1.35 ml tubes: for 2 x 100-well ring discs on the QIAGEN Rotor-Gene Q or 250 x 20 µl reactions
¥37,000
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RT² SYBR Green ROX FAST Mastermix (12)

Cat. No. / ID:  330622

Contains 12 x 1.35 ml tubes: for 12 x 100-well ring discs on the QIAGEN Rotor-Gene Q or 1500 x 20 µl reactions
¥199,000
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RT² SYBR Green FAST Mastermix (24)

Cat. No. / ID:  330603

Contains 24 x 1.35 ml tubes: for 24 x 100-well ring discs on the QIAGEN Rotor-Gene Q or 3000 x 20 µl reactions
¥310,000

特徴

  • 最小限のプライマーダイマー形成で特異的な増幅
  • 装置に特化したオールインワンのマスターミックス
  • ROX、Fluoresceinまたは色素無しのマスターミックスが入手可能
  • 通常のSYBR Greenマスターミックスよりも短時間のプロトコール
  • 高感度で明確な解離曲線

製品詳細

RT2 SYBR Green FAST MastermixesはSYBR Greenを用いたリアルタイムPCRアプリケーション(reference dye添加/未添加)に最適です。RT2 SYBR Green FAST Mastermixesは感度や特異性に影響を与えずに迅速なPCRプロトコールを使用するために最適化されています。RT2 SYBR Green FAST MastermixesはRT2 qPCR Primer AssayおよびRT2 Profiler PCR Arrayと組み合わせてご使用ください。

パフォーマンス

RT² SYBR Green FAST Mastermixesには、RT2 qPCR Primer AssayおよびRT2 Profiler PCR Arrayを用いたSYBR GreenベースのリアルタイムPCRに必要な至適化済み試薬とバッファーがすべて入っていて、ほとんどのリアルタイムPCR装置で使用できます。

RT² SYBR Green FAST Mastermixesを用いて行なった高性能リアルタイムPCRは、高い増幅効率(図“ 幅広いダイナミックレンジで高い増幅効率”)および高レベルの感度と特異性(図“ ポリメラーゼ活性の厳密な制御により高い特異性を実現”)を実証しています。

さらにRT2 SYBR Green FAST Mastermixesを用いて得られた解離曲線は、通常のサイクリング条件で得られる解離曲線に比べてバックグランドが低く、より明確な結果が得られます(図“ RT2 SYBR Green Fast Mastermixを用いて明確な解離曲線を実現”)。

図参照

原理

RT2 SYBR Green FAST Mastermixesには、リアルタイムPCRバッファー、高性能HotStart DNA Taq polymerase、ヌクレオチド、SYBR Green色素が入っています。装置の光学系を最適化するために、フルオレセインまたはROX色素のいずれかが入ったマスターミックスもあります。化学修飾され厳密にコントロールされたHotStart酵素を用いることにより、プライマーダイマーあるいは非特異的な産物の増幅を回避でき、正確なSYBR Green結果が得られます。RT2 SYBR Green FAST Mastermixesは、10秒間の変性ステップ、30秒間のアニーリング/エクステンションステップをもつPCRプロトコール用に至適化され、特異性や感度の低下はありません。

ROXあるいはフルオレセインを含まないマスターミックス

RT2 SYBR Green FAST Mastermixは reference dyeが不要な装置でのSYBR Greenベースの検出用リアルタイムPCRアプリケーションに最適です。 対象装置: Bio-Rad models CFX96、CFX384; Bio-Rad/MJ Research Chromo4、DNA Engine Opticon、DNA Engine Opticon 2; Roche LightCycler 480 (96-well and 384-well); Eppendorf Mastercycler ep realplex without ROX filter set; Cepheid SmartCycler。

フルオレセインを含むマスターミックス

RT2 SYBR Green Fluor FAST Mastermixは、 reference dyeとしてフルオレセインを使用する装置でのSYBR Greenベースの検出用リアルタイムPCRアプリケーションに最適です。 対象装置:Bio-Rad models iCycler、iQ5、MyiQ、MyiQ2。

ROXを含むマスターミックス

RT2 SYBR Green ROX FAST Mastermixは、 reference dyeとしてROXを使用する装置でのSYBR Greenベースの検出用アルタイムPCRアプリケーションに最適です。 対象装置:QIAGENの Rotor-Gene Q; Applied Biosystems models 5700, 7000, 7300, 7500 (Standard and Fast), 7700, 7900HT (Standard and Fast 96-well block, 384-well block), StepOnePlus, ViiA 7 (Standard and Fast 96-well block, 384-well block); Eppendorf Mastercycler ep realplex with or without ROX filter set; Stratagene models Mx3000P, Mx3005P, Mx4000;Takara TP-800。

アプリケーション

RT2 SYBR Green FAST MastermixesはリアルタイムPCRアプリケーションでの使用に最適です。

裏付けデータと数値

リソース

MSDS (1)
Download Safety Data Sheets for QIAGEN product components.
キットハンドブック (1)
For pathway-focused gene expression profiling using real-time RT-PCR
Safety Data Sheets (1)
Certificates of Analysis (1)

FAQ

Are primers available that only detect mitochondrial DNA encoded genes and not nuclear genomic DNA encoded genes?
There are less than a dozen genes encoded by the mitochondrial genome (all other mitochondrial proteins are encoded by nuclear genes), and they are all transcribed as one transcript (just like any prokaryote), so distinguishing the expression of individual genes by real-time RT-PCR is not possible.
FAQ ID -2680
How can I ensure that reaction volume is not lost due to evaporation during thermal cycling?
Be sure to carefully and completely seal the qPCR assay plate with fresh, optical, thin-wall, 8-cap strips or adhesive optical film before the plate is placed into the real-time cycler. In addition, refer to your instrument's user's manual to determine whether the real-time cycler manufacturer recommends use of a plate compression pad during the run.
FAQ ID -2679
How do I create a workspace that is free of DNA contamination, prior to carrying out a qPCR experiment?

Any DNA contamination will artificially inflate the SYBR Green signal, yielding skewed gene expression profiles and false-positive signals. The most common source of DNA contamination is from PCR products generated during previous experiments. Such contamination is most often due to the improper disposal of tubes, tips, and gels that previously came into contact with PCR products. Additionally, PCR products may also contaminate pipettors, racks, work pads, and commonly used reagents such as water and buffers. To minimize the risk of contaminating your experiment with extraneous DNA, the following steps should be taken:

 

  • Remove a single aliquot of water from your PCR-grade stock, sufficient to complete the experiment. This minimizes the number of times that the stock container is opened, thereby minimizing contamination risks.
  • Use only fresh PCR-grade reagents and disposable labware.
  • Treat any labware (tubes, tips, and tip boxes) used in PCR with 10% bleach, before discarding.
  • Maintain a dedicated workspace for PCR setup (perhaps a PCR-only hood), away from areas of the lab where post-PCR work is done, such as running gels, enzyme digestions, and cloning.
  • Change the lab bench pads/papers often and decontaminate lab benches and labware (racks, pipettors, etc.) before each use by washing with 10% bleach, and/or exposing to UV light for at least 10 minutes. This serves to degrade and/or inactivate contaminating DNA.
  • Before, during, and after the experiment, minimize the opening and closing of any tubes or plates used during the experiment.  
FAQ ID -2654
What is a dissociation curve, and why is it important to run a dissociation curve, following qPCR using SYBR Green chemistry?

Dissociation curves are carried out at the end of a PCR experiment by following a 3-step procedure.

First, all the components are denatured at 95°C, followed by complete annealing at a set temperature (based on the primer Tm values), followed by a gradual increase in temperature up to 95°C. Fluorescence intensity is monitored during this final temperature increase, resulting in the generation of a melting curve or dissociation curve.

By analyzing the first derivative of such a curve, you can readily assess the homogeneity of the PCR products, including the presence of primer–dimers, thereby determining the specificity of the PCR reaction. It is important to carry out such post-PCR analyses when using SYBR Green probe chemistry due to this reagent's lack of sequence specificity.

FAQ ID -2678