QIAwave Miniprep Kit – Eco-friendlier Plasmid Extraction

Alternative plus respectueuse de l’environnement que notre kit standard pour extraire jusqu’à 20 µg d’ADN plasmidique de qualité biologie moléculaire

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Contactez notre équipe et demandez un devis pour un kit d’essai QIAwave Plasmid Miniprep Kit (50).

QIAwave Plasmid Miniprep Kit (50)

Cat. No. / ID:   27204

QIAprep 2.0 Spin Columns, Waste Tubes (2 ml), Reagents  
125,00 $US
Connexion Pour ouvrir votre compte.
KitKit component
Eco-friendlier kit
Eco-friendlier Collection Tubes
Preparations
50
250
Le QIAwave Plasmid Miniprep Kit (50) est destiné aux applications de biologie moléculaire. Ce produit n’est pas conçu pour le diagnostic, la prévention ou le traitement des maladies.
Vous aimeriez essayer cette solution ?
Contactez notre équipe et demandez un devis pour un kit d’essai QIAwave Plasmid Miniprep Kit (50).

Caractéristiques

  • Qualité et performance de l’ADN plasmidique identiques à celles du QIAprep Spin Miniprep Kit
  • Jusqu’à 22 % de plastique en moins et jusqu’à 14 % de carton en moins par rapport au QIAprep Spin Miniprep Kit
  • Tubes de décharge réutilisables constitués à 100 % de plastique recyclé post-consommation
  • Les tampon concentrés utilisent jusqu’à 93 % de plastique en moins que nos tampons standard

 

Détails produit

Le QIAwave Plasmid Miniprep Kit est une version plus respectueuse de l’environnement que notre kit standard QIAprep Spin Miniprep Kit. Le QIAwave Kit utilise jusqu’à 22 % de plastique en moins et jusqu’à 14 % de carton en moins que notre kit standard, et propose des tubes de décharge constitués à 100 % de plastique recyclé post-consommation qui peuvent être réutilisés tout au long de la procédure. Les tampons QIAwave se présentent sous une forme concentrée, réduisant ainsi de 93 % la quantité de plastique par flacon. Pour économiser du papier, aucun protocole imprimé n’a été inclus dans le kit. Vous pouvez télécharger les protocoles à partir de la liste des ressources ou scanner le code QR situé à l’intérieur du kit. Bien que l’emballage et les composants de notre kit QIAwave puissent sembler différents, il sont aussi faciles à utiliser que notre kit standard, et la chimie et la performance sont identiques.


Veuillez noter que vous aurez besoin de flacons en verre stériles pour stocker les tampons reconstitués.


En partenariat avec My Green Lab, nous avons également évalué l’impact environnemental de ce kit. Les labels My Green Lab ACT sont destinés à évaluer et à noter les produits selon plusieurs critères de durabilité. Ceux-ci incluent :


• Fabrication
• Gestion responsable des produits chimiques
• Contenu durable dans les produits et les matériaux d’emballage
• Élimination de l’emballage à la fin de sa vie utile

Les produits sont notés de 1 à 10, à l’exception des produits de consommation d’eau et d’énergie à qui l’on attribue 1 point par kWh ou gallon, respectivement. Un faible score signifie un impact environnemental plus faible – voir les figures « Label du facteur d’impact environnemental du QIAwave Plasmid Miniprep Kit ACT US  50/ 250, EU  50/ 250 et UK 50/ 250 ».

Le QIAwave Plasmid Miniprep Kit est conçu pour isoler jusqu’à 20 μg d’ADN plasmidique ou de cosmide très pur pour les applications de biologie moléculaire de routine, notamment le séquençage et le clonage fluorescents et radioactifs. Vous pouvez obtenir des rendements encore plus élevés (jusqu’à 30 μg) à l’aide du Protocole complémentaire à haut rendement QIAprep. Pour des résultats optimaux, nous vous recommandons d’utiliser ce kit avec le QIAvac 24 Plus.

 

Voir les illustrations

Performances

Les performances entre nos QIAwave Plasmid Miniprep Kit et QIAprep Spin Miniprep Kit sont identiques car la chimie est la même. Nous avons également démontré que les deux kits sont plus performants que les kits concurrents (voir la figure « Performance du kit QIAwave »).

Le QIAwave Plasmid Miniprep Kit vous permet de purifier jusqu’à 20 μg d’ADN plasmidique ou de cosmide de qualité biologie moléculaire pour les applications de biologie moléculaire de routine telles que la PCR, le séquençage et le clonage.

Les QIAprep 2.0 Spin Columns sont si polyvalentes que vous pouvez les utiliser dans des microcentrifugeuses, sur des collecteurs de vide ou sur le QIAcube Connect (voir les figures « Options de manipulation des QIAprep 2.0 Spin Columns :  microcentrifugeuse,  collecteurs de vide et  système automatisé »). La procédure sous vide simplifie la manipulation et accélère le traitement des échantillons. Les QIAprep 2.0 Spin Columns peuvent également être traitées sous vide avec le QIAvac 24 Plus ou tout autre collecteur commercial avec des raccords luer.

Format Colonnes de centrifugation
Module de purification QIAprep 2.0 Spin Columns
Débit 1 à 24 échantillons
Temps de préparation 24 minipréparations en 30 minutes
Équipement requis Microcentrifugeuse ou collecteur de vide ; peut être entièrement automatisé avec le QIAcube Connect
Nettoyage des lysats Centrifugation
Capacité du réservoir de la colonne 800 µl
Volume minimum du tampon d’élution 50 µl
Volume de culture pour les plasmides à copies élevées 1 à 5 ml
Volume de culture pour les plasmides/cosmides à copies faibles 1 à 10 ml

L’ADN purifié peut être utilisé dans la digestion de restriction (voir la figure «  Digestion complète avec diverses enzymes de restriction »).

Nous avons également comparé les rendements d’ADN plasmidique obtenus à l’aide du tampon QIAwave Plasmid Miniprep Kit (50), préparé par versement ou pipettage, et des tampons standard QIAprep Spin Miniprep Kit (50). Les deux méthodes ont permis l’obtention de rendements comparables comme le montre la figure «  Manipulation des tampons concentrés ».

 

Voir les illustrations

Principe

Les QIAprep 2.0 Spin Columns contiennent une membrane de silice unique qui lie jusqu’à 20 µg d’ADN en présence d’une forte concentration de sels chaotropiques et permet l’élution dans un faible volume de tampon faiblement salin. La technologie de membrane QIAprep élimine la longue extraction de phénol-chloroforme et la précipitation d’alcool, ainsi que les problèmes et tracas liés aux résines et coulis libres. L’ADN plasmidique de pureté élevée élué dans les QIAprep 2.0 Spin Columns est prêt à l’emploi – inutile de précipiter, concentrer ou dessaler.

 

Procédure

La purification de l’ADN plasmidique avec QIAwave Plasmid Miniprep suit une procédure simple de liaison-lavage-élution (voir le schéma «  Procédure QIAwave plasmid Miniprep »).

1. Lysez les cultures bactériennes et nettoyez les lysats par centrifugation.

2. Ajoutez les lysats nettoyés aux QIAprep 2.0 Spin Columns. À ce stade, l’ADN plasmidique est adsorbé dans la membrane de silice et les impuretés sont éliminées.

3. L’ADN pur est ensuite élué dans un faible volume de tampon d’élution ou d’eau.

En plus de la purification de l’ADN plasmidique d’E. coli, le QIAwave Plasmid Mini Kit peut être utilisé pour purifier l’ADN plasmidique de Saccharomyces cerevisiae, Bacillus subtilis et Agrobacterium tumefaciens. Contactez notre équipe de services techniques ou votre distributeur local si vous avez besoin de protocoles pour ces applications.

Les tampons QIAwave se présentent sous une forme concentrée pour une reconstitution facile par ajout d’eau et/ou d’éthanol ; veuillez consulter le manuel pour plus de détails. Les QIAwave QIAprep 2.0 Spin Columns et QIAwave QIAprep 2.0 Waste Tubes sont conditionnés en sachets individuels et doivent être préassemblés avant le début du protocole. Cela prend un peu de temps, mais réduit les déchets plastiques.

Le QIAwave Plasmid Miniprep Kit peut être automatisé sur le QIAcube Connect à l’aide des protocoles du QIAprep Spin Miniprep Kit.

 

Voir les illustrations

Applications

Le QIAwave Plasmid Miniprep Kit fournit des rendements reproductibles d’ADN de pureté élevée adaptés à la plupart des applications, y compris :

  • PCR
  • Digestion par enzymes de restriction
  • Ligature et transformation
  • Séquençage
  • Dépistage

 

Données et illustrations utiles

Specifications

FeaturesSpecifications
ApplicationsSéquençage Séquençage fluorescent et radioactif (y compris le séquençage capillaire), la ligature, le clonage, la transformation, etc.
ProcessingManual (centrifugation or vacuum)
Plasmid typeHigh-copy, low-copy, cosmid DNA
Culture volume/starting material1–10 mL culture volume
Elution volume50 µL (minimal)
TechnologySilica technology
Time per run or prep per run<30 minutes
Yield<20 ug
Samples per run (throughput)1–24 samples per run
Number of preps per run1–24 samples per run

Ressources

Application/Protocol Documents (1)
Step up your sustainability by recycling your labware. This handy guide will show you how to quickly and easily recycle kit components and reduce plastic waste in your lab.
Brochures & Guides (2)
Quick-Start Protocols (1)
Kit Handbooks (1)
Safety Data Sheets (1)
Certificates of Analysis (1)

FAQ

What is the advantage of running an analytical gel with fractions of my plasmid preparation?

Running fractions saved from each step in the plasmid preparation procedure on an agarose gel enables monitoring the performance of each crucial step in the protocol. If the plasmid DNA is of low yield or quality, the samples can be analyzed to determine at what stage of the purification procedure the difficulty occurred.

Aliquots can be taken from the cleared lysate and the flow-throughs as indicated in the relevant protocols, precipitated with isopropanol and resuspended in a small volume of TE buffer.

Please see the Troubleshooting Section of the QIAprep Miniprep Handbook and Appendix A of the QIAGEN Plasmid Purification Handbook for instructions, and a picture and legend explaining the typical results you may see. You can also access this information on our Plasmid Resource Pages.

FAQ ID -769
I don’t have any glassware in the lab, is the QIAwave kit still a good option for me?
The concept of QIAwave includes the reconstitution of functional buffers. We recommend the use of glass bottles for this procedure. Glass bottles are easier to clean, sterilize and reuse than plastic bottles, further reducing the plastic footprint of the kit. If you do not have the option of using clean glass or plastic bottles, we recommend using our legacy kits. 
FAQ ID - 3989

What is the recipe for LB?
Preparation of LB medium: Dissolve 10 g tryptone, 5 g yeast extract, and 10 g NaCl in 800 ml dH2O. Adjust the pH to 7.0 with 1 N NaOH. Adjust the volume to 1 liter with dH2O. Sterilize by autoclaving.
FAQ ID -212
How do I know if my plasmid is a high- or low copy number type?

Find out which origin of replication your plasmid contains, and look at the table below for classification into high-copy or low-copy types. This table can also be found online at the QIAGEN Plasmid Resource Center in the section 'Growth of bacterial cultures; Plasmid Copy Number' . A way to determine experimentally if the copy number of your plasmid is high or low is to perform a miniprep. A high-copy plasmid should yield between 3-5 ug DNA per 1 ml LB culture, while a low-copy plasmid will yield between 0.2-1 ug DNA per ml of LB culture.

 

Origins of replication and copy numbers of various plasmids and cosmids

DNA construct Origin of Replication Copy number Classification
Plasmids      
pUC vectors pMB1* 500–700 high copy
pBluescript® vectors ColE1 300–500 high copy
pGEM® vectors pMB1* 300–400 high copy
pTZ vectors pMB1* >1000 high copy
pBR322 and derivatives pMB1* 15–20 low copy
pACYC and derivatives p15A 10–12 low copy
pSC101 and derivatives pSC101 ~5 very low copy
Cosmids      
SuperCos pMB1* 10-20 low copy
pWE15 ColE1 10-20 low copy

* The pMB1 origin of replication is closely related to that of ColE1 and falls in the same incompatibility group. The high-copy plasmids listed here contain mutated versions of this origin.

FAQ ID -350
Can I eliminate RNase A from buffer P1 for my plasmid preparation to obtain RNase-free DNA for in-vitro transcription?
No, RNase A should not be omitted from buffer P1. It is required to prevent RNA contamination of the purified plasmid DNA. RNase A will not interfere with downstream in-vitro transcription experiments, since it will be efficiently removed during the plasmid purification procedures using QIAGEN Plasmid Kits.
FAQ ID -366
I left Buffer P1 at room temperature after addition of RNase A, what shall I do?

Buffer P1 with RNase A used in QIAGEN Plasmid Purification Kits should be fine at room temperature for a few days. We would expect the enzyme to have some residual activity. However, optimal results cannot be guaranteed after storage at room temperature. If you notice that RNase A activity is substantially reduced, you can add fresh RNase A to your buffer.

We recommend that Buffer P1 with RNase A be stored in the refrigerator (2–8°C). RNase A will be stable for 6 months under this condition.

FAQ ID -859
Can the QIAwave kits be recycled?
We provided an infographic describing the composition of most of our purification kits. You can use the information provided as a guide to recycle kit components and use it to reduce plastic waste in your laboratory. Depending on the specific kit and application, certain kit components may contain or come into contact with chemicals and biological samples, in this case, the components should be disposed of according to local guidelines and regulations. You can find more information on More Sustainable Products (qiagen.com).
FAQ ID - 3992
What water should I use to prepare the buffer concentrates?
We recommend using highly pure water for reconstitution. Ultrapure water (such as the one from MilliQ system, also known as type 1 water) with a resistivity of 18.2 MΩ-cm at 25°C can be used. In case you do not have access to type 1 water, QIAGEN offers Nuclease-Free Water (5 L, cat. No. 129117); and Nuclease-Free Water (1000 mL, cat. No. 129115). It is important that you do not use tap water as this may interfere with the extraction of the target analyte. 
FAQ ID - 3986

How do I perform a DNA precipitation to concentrate my sample?
  • Add 1/10 volume of 3 M Na-Acetate pH 5.2, and 2 to 2.5 volumes of ice-cold 100% ethanol to the DNA sample
  • Mix, and store at –20°C for at least 1 h to precipitate the DNA
  • Recover the precipitated DNA by centrifugation at full speed in a microcentrifuge for 15–20 min
  • Pour off the ethanol and wash the pellet twice with room-temperature 70% ethanol
  • Allow the DNA pellet to air-dry
  • resuspend the DNA in a suitable volume of sterile TE buffer or distilled water
FAQ ID -305
With LyseBlue reagent for lysis control, can I now process more bacterial culture and overload the columns?

No. The maximum culture volumes recommended for QIAGEN's plasmid preparation kits still apply, and should be strictly followed. LyseBlue reagent now allows the user to monitor potential problems (insufficient bacterial cell resuspension and lysis as a consequence of overloading) early in the plasmid preparation process.

FAQ ID -864
What is the recipe for 2x YT?
2x YT medium, per liter 16 g tryptone 10 g yeast extract 5 g NaCl Media Preparation and Bacteriological Tools. Edited by: Fred M. Ausubel, Roger Brent, Robert E. Kingston, David D. Moore, J.G. Seidman, John A. Smith, Kevin Struhl Current Protocols in Molecular Biology (1994), Section 1.1.3.
FAQ ID -213
What are the additional plasmid bands I see on my gel?

Open circular plasmid, resulting from single strand nicks, usually migrates slower in agarose gels and forms (faint) bands above the supercoiled plasmid DNA band. Sometimes an additional band of denatured supercoiled DNA migrates just below the supercoiled form. This form may result from prolonged alkaline lysis with Buffer P2 and is resistant to restriction digestion.

For a detailed description on how to run and interpret an analytical gel, please see Appendix A in the QIAGEN Plasmid Purification Handbook: "Agarose Gel Analysis of the Purification Procedure", or visit this link.

FAQ ID -1059
Do you have a protocol for the isolation of plasmid DNA from Bacillus subtilis?
Is there a way to compare the environmental impacts of the kit?
In partnership with My Green Lab, we were able to assess the environmental impact of the kits. My Green Lab ACT (accountability, consistency and transparency) environmental impact factor labels are designed to evaluate and score products on several sustainability criteria. The products are scored from 1 to 10, except for energy and water consumption which are scored as 1 point per kWh or gallon, respectively. A low score means a lower environmental factor.  
FAQ ID - 3991

What is the composition of Buffer PB?
Buffer PB contains a high concentration of guanidine hydrochloride and isopropanol. The exact composition of Buffer PB is confidential. However, this buffer can be purchased separately: Buffer PB.
FAQ ID -2791
Does the reuse of the Waste Tubes increase the risk of cross-contamination?
No, we were able to prove experimentally that cross-contamination does not occur through the reuse of the Waste Tubes. The Waste Tubes are used to collect the flow-through from the lysis and washing steps, where the flow-through is discarded afterwards. If the flow-through is to be used for further processing, we recommend the use of a Collection Tube
FAQ ID - 3988

Do you have a protocol for obtaining > 20 µg plasmid DNA using the QIAprep Spin Miniprep Kit?
Why is my plasmid DNA yield low?

Low yields of plasmid DNA can be caused by a number of different factors. The most common causes for low yield are poor culturing conditions and plasmid propagation, excessive amounts of starting material resulting in insufficient bacterial cell lysis and column overloading. When working with the anion-exchange based QIAGEN Plasmid Purification Kits, extra care is required during the isopropanol precipitation step, as the glassy DNA pellet may be difficult to see, and tends to be only loosely attached to the side of the tube.

We strongly recommend to review the information provided on our Plasmid Resource Page in the section 'Optimal results with QIAGEN plasmid kits', as it provides useful background information on growing bacterial cultures and general considerations for optimal results. It is also necessary to follow the instructions in the relevant protocols precisely to ensure the best plasmid yield and quality.

To determine at what stage of the procedure any problem occurred, save fractions from different steps of the purification procedure, and analyze by agarose gel electrophoresis. For a detailed description on how to run and interpret an analytical gel, please see Appendix A in the QIAGEN Plasmid Purification Handbook: "Agarose Gel Analysis of the Purification Procedure", or visit the QIAGEN Plasmid Resource Center.

 

 

FAQ ID -768
Are the QIAwave kits based on a different chemistry than the legacy kits?
No, the chemistry between the QIAwave kits and the legacy kits is the same, and so is the performance. The QIAwave buffers come as concentrates, reducing the amount of plastic per bottle while maintaining the same functionality after reconstitution. The advantage of the QIAwave kits is the reduction of materials used for the kits, such as plastics, cardboard and paper. In addition, the QIAwave kits contain Waste Tubes made from 100% post-consumer plastic. 
FAQ ID - 3990

Why do I get genomic DNA contamination in my plasmid prep?

Too vigorous mixing of the bacterial lysate causes genomic DNA to appear in the eluate. The lysate must be handled gently after addition of buffers P2 and P3 to prevent shearing of chromosomal DNA. The culture volume needs to be reduced if the lysate is too viscous for gentle mixing.

Use of LyseBlue Reagent enables visualization of efficient bacterial cell resuspension as a prerequisite for complete lysis, thereby helping to avoid overloading of the columns and additional difficulties related to highly viscous lysates.

Additional information for successful plasmid preparations using QIAGEN's broad selection of Plasmid Kits can be found at our Plasmid Resource Center.

FAQ ID -353
Do you have a protocol for the isolation of plasmid DNA from Agrobacterium?
FAQ ID -898
I am seeing a precipitate after adding LyseBlue reagent to Buffer P1. What should I do about that?

A precipitate forming upon adding LyseBlue reagent to Buffer P1 is a normal observation. This precipitate will completely dissolve after addition of Buffer P2. Please be sure to shake Buffer P1 vigorously before use to completely resuspend LyseBlue particles.

FAQ ID -1045
What to do if cell clumps are present after Buffer P2 addition when using LyseBlue Reagent?

If cell clumps are present after adding Buffer P2 to your samples when using a QIAGEN Plasmid Purification Kit in combination with LyseBlue Reagent, you have two options:

  • pipet the cell clumps up and down for resuspension
  • transfer any clumps to a separate tube, add Buffer P1 and mix vigorously for resuspension, add Buffer P2 for lysis, and subsequently transfer the lysed material back to combine it with the rest of the original solution

Note: Avoid incubation times longer than 5 minutes in Buffer P2 as this will irreversibly denature plasmid DNA. In the scenario above, Buffer P3 may need to be added to portions of the sample, which can be subsequently combined once resuspension, lysis and neutralization of all fractions is complete.

FAQ ID -861
What is the new Waste Tube made of?
The new Waste Tube is made from recycled plastic recovered from post-consumer plastic waste. Due to the slight composition differences of the raw material, the color of the tubes may differ from lot to lot. However, this has no effect on its intended use of collecting flow-through from sample binding and membrane washing. After each step, the flow-through is discarded, and the Waste Tube can be reused. The Waste Tube is only used to process waste and should never come into direct contact with the analyte of interest. For detailed instructions, you can watch our instructional video at www.qiagen.com/qiawavewastetube
FAQ ID - 3987

What is the recipe for SOC medium?

The components of the SOC medium are:

  • 0.5% Yeast Extract
  • 2% Tryptone
  • 10 mM NaCl
  • 2.5 mM KCl
  • 10 mM MgCl2
  • 10 mM MgSO4
  • 20 mM Glucose*

*Note: add Glucose after autoclaving the solution with the remaining ingredients, and letting it cool down. Sterilize the final solution by passing it through a 0.2 µm filter.

SOC medium can be stored at room temperature and is stable for several years.

FAQ ID -798