QIAGEN Plasmid Plus Kits

最高10 mgまでのトランスフェクショングレードのプラスミドDNAを最も迅速かつ簡便に精製

S_1084_5_GEN_V2
Want to try this solution for the first time?
Get in touch with our team today and request a quote for your QIAGEN Plasmid Plus Midi Sample Kit (5) trial kit.
QIAGEN Plasmid Plus Midi Sample Kit (5)

Cat. No. / ID:  12941

QIAGEN Plasmid Plus Midi Sample Kit (5)
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KitAccessories
QIAGEN Plasmid Plus Kit
QIAvac Holder
VacValves
Column type
Midi
Maxi
Mega
Giga
Preparations
5
25
100
QIAGEN Plasmid Plus Kits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。
Want to try this solution for the first time?
Get in touch with our team today and request a quote for your QIAGEN Plasmid Plus Midi Sample Kit (5) trial kit.

Features

  • 最大24サンプルを迅速に精製
  • 迅速な吸引操作で同時調製可能
  • 少ない溶出量で高濃度DNA
  • 特殊な洗浄バッファーで非常に低いエンドトキシンレベル(<1 EU/µg)

Product Details

QIAGEN Plasmid Plus Kits は最高10 mgの高純度プラスミドDNAをラージスケールで非常に迅速に精製します。吸引マニホールドを用いて最高24個のサンプルを同時に精製でき、マニュアルでの作業時間を短縮できます。溶出量が少ないので高濃度のプラスミドDNAが得られ、すぐに使用できます。 エタノール沈殿は不要です。さらに QIAGEN Plasmid Plus Kits には、洗浄バッファーが含まれており、精製されたプラスミドはエンドトキシンレベルが低いという特長もあります。 精製されたプラスミドは様々なアプリケーションに最適で、感受性の高い細胞株にもトランスフェクトできます。

Performance

安定した品質および性能

QIAGEN Plasmid Plus テクノロジーは、陰イオン交換テクノロジーと同等の性能および純度を実現します。QIAGEN Plasmid Plus columns のユニークな結合ケミストリーにより高純度なプラスミドDNAが安定して精製できます。高収量プロトコールおよびそれに必要な追加のバッファー量もIAGEN Plasmid Plus Kits に入っており、RNA、タンパク質、ゲノムDNAなどの夾雑物を含まない高純度のプラスミドDNAを最高10 mg(Giga)、2 mg(Mega)、1 mg(Maxi)、250 µg(Midi)まで得られます(図 " QIAGEN Plasmid Plus Kits(QPP)は高収量を実現"  および " 分析クロマトグラム")。

エンドトキシンレベルが非常に低いトランスフェクショングレードDNA

リポ多糖類またはLPSとして知られているエンドトキシンは、E. coli などのグラム陰性菌の細胞膜成分であり、プラスミド精製の溶解ステップ中に放出されます。エンドトキシンは非特異的な免疫応答の活性化を誘導するため、トランスフェクション実験のセットアップにおいてコントロールできない変動要素となり、実験結果およびその再現性に影響し、結果の比較および解釈が困難になります。  

キットに添付のエンドトキシン洗浄バッファーを利用することにより、バクテリア由来のエンドトキシンレベルを通常1 EU/μg DNA 以下に減らすことができます。エンドトキシンレベルは宿主菌株、培養液、細胞密度により変動します。細胞密度が非常に高い場合には、エンドトキシンレベルが13 EU/μg DNA の範囲で多少高くなることがあります。このような特性により、精製したプラスミドDNAは感受性の高い細胞株へのトランスフェクションをはじめとする多くのアプリケーションに最適です (図 " QIAGEN Plasmid Plus Kits を用いた場合のエンドトキシンレベル"、" 様々なプラスミドDNA 調製法を用いた際のエンドトキシンレベル"、" 感受性の高い細胞株への効果的なトランスフェクション")。 

See figures

Principle

QIAGEN Plasmid Plus Kitsはラージスケールのプラスミドを簡単に調製するため方法です。QIAGEN Plasmid Plus columnsのデザインおよびユニークな結合ケミストリーにより、結合、洗浄、溶出ステップが簡便に行なえます。この操作によりQIAGEN Plasmid Plus Kits は超高速なラージスケールのプラスミド調製を実現し、MaxiおよびMidi調製にかかる時間は20 分です。得られた高濃度なプラスミドDNAは、続くアプリケーションにすぐに使用できます。エタノール沈殿は不要です。このように、本キットで得られるプラスミドDNAは、微量の反応容量を必要とするアプリケーションに最適です。

LyseBlueを用いて最大のプラスミド収量を実現

バッファーの混和が最適に行なわれているかを簡単に目視で判断できる、オプションの色素指示薬LyseBlueもキットに含まれています。この試薬により、非効率的な溶菌操作およびSDS、ゲノムDNA、細胞残渣の不完全な沈殿を引き起こす一般的な操作ミスを防ぐことができます。このユニークなバッファー添加剤は、溶液の色変化だけで最適な溶菌操作および溶液の中和を確認でき、収量の増大を実現します。

製品仕様

特徴
Plasmid Plus
Giga Kit
Plasmid Plus 
Mega Kit
Plasmid Plus 
Maxi Kit
Plasmid Plus 
Midi Kit
Applications Transfection of most cell lines, cloning, enzymatic restriction,
in-vitro translation/transcription, sequencing
Transfection of most cell lines, cloning, enzymatic restriction,
in-vitro translation/transcription, sequencing
Transfection of most cell lines, cloning, enzymatic restriction,
in-vitro translation/transcription, sequencing
Transfection of most cell lines, cloning, enzymatic restriction,
in-vitro translation/transcription, sequencing
Culture volume/starting material 2500 ml culture volume 500 ml culture volume 50–200 ml culture volume 25–50 ml culture volume
Final DNA concentration ≥1 µg/µl ≥1 µg/µl ≥1 µg/µl ≥1 µg/µl
Plasmid type Plasmids, cosmids Plasmids, cosmids Plasmids, cosmids Plasmids, cosmids
Processing Manual (centrifugation and vacuum), parallel processing Manual (centrifugation and vacuum), parallel processing Manual (centrifugation and vacuum), parallel processing Manual (centrifugation and vacuum), parallel processing
Samples per run; throughput 1–12 samples per run 1–12 samples per run 1–24 samples per run 1–24 samples per run
Technology Advanced silica technology (non-chaotrophic), endotoxin reduction Advanced silica technology (non-chaotrophic), endotoxin reduction Advanced silica technology (non-chaotrophic), endotoxin reduction Advanced silica technology (non-chaotrophic), endotoxin reduction
Time per run 50 min 40 min  20 min 20 min
Yield up to 10 mg up to 2.5 mg up to 1 mg up to 250 µg

Procedure

QIAGEN Plasmid Plus Kits に添付のQIAfilter Cartridges によりライセートを迅速に清澄化できます。1ステップで清澄化ライセートをカラムにアプライできるので時間の節約に繋がります。QIAGEN Plasmid Plus columnにアプライした清澄化ライセートおよび続いて行なう洗浄バッファーの吸引には吸引マニホールド(QIAvac 24 Plus)を使用します。メンブレンの洗浄および乾燥後、遠心操作により高濃度DNAが少量で溶出されます(図 " QIAGEN Plasmid Plus 操作手順")。
See figures

Applications

QIAGEN Plasmid Plus Kitsは以下のようなアプリケーションに最適なトランスフェクショングレードのプラスミドDNA を調製します。

  • Huh-7のような感受性の高い細胞株を含むほとんどの細胞株へのトランスフェクション
  • 制限酵素分解
  • クローニング
  • In vitro 転写/トランスレーション
  • シークエンシング

Supporting data and figures

Resources

キットハンドブック (1)
テクニカルインフォメーション (3)
Since QIAGEN launched the world’s first kit for purification of plasmid DNA, technologies have been developed to make plasmid DNA preparation even more convenient and reduce endotoxin levels in line with scientists’ needs in the laboratory. A lot of well-known companies have developed kits, and the choice of solutions available has never been greater. We compared popular advanced-technology kits to see their relative performance in speed, convenience of handling, yield and endotoxin levels.
MSDS (2)
Download Safety Data Sheets for QIAGEN product components.
Download Safety Data Sheets for QIAGEN product components.
クイックスタートプロトコール (3)
パンフレット (1)
Certificates of Analysis (1)
Brochures & Guides (1)
Technical Information (3)
Since QIAGEN launched the world’s first kit for purification of plasmid DNA, technologies have been developed to make plasmid DNA preparation even more convenient and reduce endotoxin levels in line with scientists’ needs in the laboratory. A lot of well-known companies have developed kits, and the choice of solutions available has never been greater. We compared popular advanced-technology kits to see their relative performance in speed, convenience of handling, yield and endotoxin levels.

FAQ

Can I use your QIAvac 96 for extraction of the DNA using QIAamp 96 DNA Blood Kit?

We do not recommend using a vacuum manifold for DNA extraction using the QIAamp 96 DNA Blood Kit because:

  1. Viscosity of the lysates varies from sample to sample. Therefore, the wells of the QIAamp 96 plate are at great risk of clogging.
  2. Use of vacuum manifold would result in incomplete removal of ethanol and salts after washing with Buffer AW2. This will affect the purity and yield of samples.
FAQ ID -9158
How can I purchase replacement bottles, caps and tubing for my QIAvac Connecting System?

Please contact Nalgene for ordering. Part numbers from Nalgene:

-          10 liter waste container (Nalgene Heavy Duty Vacuum Carboy [PP], cat. no. 2226-0020)

-          20 liter waste container (Nalgene Heavy Duty Vacuum Carboy [PP], cat. no. 2226-0050)

-          Quick filling/venting closure (Nalgene, cat. no. 2158-0021) [lid]

-          Vacuum tubing (e.g., Nalgene 180 clear plastic vacuum tubing, cat. no.8000-0065)

FAQ ID -9156
Which QIAGEN plasmid preparation kits will contain LyseBlue Reagent?

LyseBlue reagent is provided in the following QIAGEN plasmid kits:

FAQ ID -865
3313 - What is the endotoxin level for the QIAGEN Plasmid Plus kits?

Endotoxin level of QIAGEN Plasmid Plus kits is in the range of 1 EU/µg DNA.

 

What is the composition of Buffer S3?
The composition of Buffer S3 is confidential. This buffer is a proprietary component of QIAGEN Plasmid Plus Kits and is an plays and important role in the alkaline lysis procedure.  Buffer S3 can only be purchased in combination with QIAGEN Plasmid Plus Kits.
FAQ ID -2788
Can the VacConnectors for your QIAvac 24 Plus be reused?

VacConnectors prevent cross-contamination and are meant to be disposed after single use.

FAQ ID -9152
Can the waste bottles of the QIAvac Connecting System be autoclaved?

Yes, the waste bottles of the QIAvac Connecting System can be autoclaved. However, please be aware that the plastic may change in color.

FAQ ID -9153
3299-Can Buffer ER from the Endofree Plasmid kits be used instead of the Buffer ETR in QIAGEN Plasmid Plus kits?

No, the two buffers cannot be interchanged.  The QIAGEN Plasmid Plus chemistry is not compatible with Buffer ER from the Endofree Plasmid kits.

Can I use your QIAvac 24 Plus for extraction of DNA from tissues using the DNeasy Blood & Tissue Kit?

We do not recommend using a vacuum manifold for DNA extraction of tissues using the DNeasy Blood & Tissue Kit. Viscosity of the lysates varies from sample to sample; therefore, the DNeasy mini spin columns are at great risk of clogging.

FAQ ID -9154
Can the QIAvac 24 Plus manifold be autoclaved?

We do not recommend to autoclave the QIAvac 24 Plus manifold. Doing so can result in damage to the device.

FAQ ID -9151
Can I use your QIAvac 96 for extraction of DNA using the DNeasy 96 Blood & Tissue Kit?

We do not recommend using a vacuum manifold for DNA extraction with the DNeasy 96 Blood & Tissue Kit because:

  1. Viscosity of the lysates varies from sample to sample. Therefore, the wells of the DNeasy 96 plate are at great risk of clogging.
  2. Use of vacuum manifold would result in incomplete removal of ethanol and salts after washing with Buffer AW2. This will affect the purity and yield of samples.
FAQ ID -9157
Is the waste tray for the QIAvac Multiwell and the QIAvac 96 the same?

QIAvac Multiwell can hold plates that conform to SBS plate standards, whereas the QIAvac 96 does not. Therefore, the waste tray for QIAvac Multiwell is not the same as for our QIAvac 96.

However, the waste tray from QIAvac Multiwell will fit the QIAvac 96, but not vice versa.

FAQ ID -9159
The gasket on my QIAvac 96 seems to have cracks in it. Do you have a replacement part available, so I can replace the gasket?
We do not have this part available for replacement. You will need to replace your vacuum manifold. Please contact your Sales representative for a price quote.
FAQ ID -9161
The gasket on my QIAvac Multiwell doesn’t seem to seal. Do you have a replacement part available, so I can replace the gasket?
FAQ ID -9160
Can I use your QIAvac 24 Plus for extraction of DNA from tissues using the DNeasy Blood & Tissue Kit?

We do not recommend using a vacuum manifold for DNA extraction of tissues using the QIAamp Blood & Tissue Kit. Viscosity of the lysates varies from sample to sample; therefore, the DNeasy mini spin columns are at great risk of clogging.

FAQ ID -9155
What is the composition of Buffer BB?
The composition of Buffer BB is confidential. Buffer BB is a proprietary component of QIAGEN Plasmid Plus Kits and functions as a binding buffer, adjusting the correct binding conditions after alkaline lysis, when performing the QIAGEN Plasmid Plus procedure. Buffer BB can only be purchased in combination with QIAGEN Plasmid Plus Kits and is not available separately.
FAQ ID -2790
What is the composition of Buffer ETR?
The composition of Buffer ETR is confidential. Buffer ETR is a proprietary component of QIAGEN Plasmid Plus Kits and functions as an endotoxin removal buffer. Buffer ETR can only be purchased in combination with QIAGEN Plasmid Plus Kits.
FAQ ID -2789
Where can I buy the filter for QIAvac Connecting System?

The filter for QIAvac Connecting System is from Pall (part no. 4400). This is the Acro 50 filter, 0.2 µm PTFE, 1/8 inch MNPT connection. Please visit www.pall.com

FAQ ID -9150