QIAwave Miniprep Kit – Eco-friendlier Plasmid Extraction

Para una alternativa más ecológica a nuestro kit estándar para extraer hasta 20 μg de ADN plasmídico de calidad de biología molecular

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QIAwave Plasmid Miniprep Kit (50)

Cat. No. / ID: 27204

QIAprep 2.0 Spin Columns, Waste Tubes (2 ml), Reagents

KitComponente del kit

Eco-friendlier kit

Eco-friendlier Collection Tubes

Preparaciones

250

El QIAwave Plasmid Miniprep Kit (50) está concebido para su uso en aplicaciones de biología molecular. Este producto no está concebido para el diagnóstico, la prevención ni el tratamiento de enfermedades.

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Características

Calidad y rendimiento del ADN plasmídico idénticos a los del QIAprep Spin Miniprep Kit
Hasta un 22 % menos de plástico y hasta un 14 % menos de cartón en comparación con el QIAprep Spin Miniprep Kit
Tubos de residuos reutilizables fabricados con plástico 100 % reciclado después de su consumo
Concentrados de soluciones tampón que utilizan hasta un 93 % menos de plástico que nuestros tampones estándar

Detalles del producto

El QIAwave Plasmid Miniprep Kit es una versión más ecológica de nuestro QIAprep Spin Miniprep Kit estándar. El QIAwave Kit utiliza hasta un 22 % menos de plástico y hasta un 14 % menos de cartón que nuestro kit estándar y ofrece tubos de residuos fabricados con plástico 100 % reciclado después de su consumo que puede reutilizar durante todo el procedimiento. Las soluciones tampón QIAwave se presentan en forma de concentrados, lo que reduce la cantidad de plástico hasta en un 93 % por botella. Para ahorrar papel, el kit no contiene ningún protocolo impreso. Puede descargar los protocolos de la lista de recursos o escanear el código QR que encontrará en el interior de la caja del kit. Aunque el envase y los componentes de nuestro QIAwave Kit pueden parecer diferentes, es tan fácil de usar como nuestro kit estándar y la composición química y el rendimiento son idénticos.

Tenga en cuenta que necesitará frascos de vidrio estériles para almacenar las soluciones tampón reconstituidas.

Además, en colaboración con My Green Lab, hemos evaluado el impacto medioambiental de este kit. Las etiquetas ACT de My Green Lab están diseñadas para evaluar y puntuar los productos según varios criterios de sostenibilidad. Entre ellos se incluyen:

• Fabricación
• Gestión responsable de los productos químicos
• Contenido sostenible en los productos y materiales de envasado
• Eliminación del envase al final de la vida útil

Los productos se puntúan de 1 a 10, excepto el consumo de energía y agua, que se puntúan con 1 punto por kWh o galón, respectivamente. Una puntuación baja implica un menor impacto medioambiental (consulte las figuras «Etiqueta del factor de impacto medioambiental del QIAwave Plasmid Miniprep Kit ACT US 50/ 250, EU 50/ 250 y UK 50/ 250»).

El QIAwave Plasmid Miniprep Kit está diseñado para aislar hasta 20 μg de ADN plasmídico o cósmido ultrapuro para aplicaciones habituales de biología molecular, que incluyen la secuenciación fluorescente y radioactiva y la clonación. Puede conseguir rendimientos aún mayores (hasta 30 μg) utilizando el QIAprep High-Yield Supplementary Protocol. Recomendamos utilizar este kit con QIAvac 24 Plus para obtener resultados óptimos.

Ver figuras

QIAwave Plasmid Miniprep Kit (50) ACT environmental impact factor label US.

QIAwave Plasmid Miniprep Kit (250) ACT environmental impact factor label US.

QIAwave Plasmid Miniprep Kit (50) ACT environmental impact factor label EU.

QIAwave Plasmid Miniprep Kit (250) ACT environmental impact factor label EU.

QIAwave Plasmid Miniprep Kit (250) ACT environmental impact factor label UK.

Rendimiento

El rendimiento entre nuestro QIAwave Plasmid Miniprep Kit y el QIAprep Spin Miniprep Kit es idéntico porque la composición química es la misma. También hemos demostrado que ambos kits superan a los de la competencia (consulte la figura « Rendimiento del QIAwave Kit»).

El QIAwave Plasmid Miniprep Kit permite purificar hasta 20 μg de ADN plasmídico o ADN cósmido de calidad de biología molecular para su uso en aplicaciones habituales de biología molecular como la PCR, la secuenciación y la clonación.

Las QIAprep 2.0 Spin Columns son tan versátiles que puede utilizarlas en microcentrifugadoras, en colectores de vacío o en el QIAcube Connect (consulte las figuras «Opciones de manejo de la QIAprep 2.0 Spin Column microcentrifugadora, colectores de vacío y sistema automatizado»). El procedimiento de vacío simplifica la manipulación y acelera el procesamiento de las muestras. Las QIAprep 2.0 Spin Columns también pueden procesarse al vacío con el uso de QIAvac 24 Plus o con cualquier otro colector con conectores luer.

Formato	Columnas de centrifugación
Módulo de purificación	QIAprep 2.0 Spin Columns
Rendimiento	1-24 muestras
Tiempo de preparación	24 minipreparaciones en 30 minutos
Equipamiento necesario	Microcentrifugadora o colector de vacío; completamente automatizables con el uso del QIAcube Connect
Aclaramiento de lisados	Centrifugación
Capacidad del depósito de la columna	800 µl
Volumen mínimo de la solución tampón de elución	50 µl
Volumen de cultivo para plásmidos de alto número de copias	1-5 ml
Volumen de cultivo para plásmidos/cósmidos de bajo número de copias	1-10 ml

El ADN purificado se puede utilizar para digestión de restricción (consulte la figura “ Digestión completa con varias enzimas de restricción”).

También hemos comparado los rendimientos de ADN plasmídico obtenidos utilizando la solución tampón QIAwave Plasmid Miniprep Kit (50), preparado por decantación o pipeteo, y las soluciones tampón estándar QIAprep Spin Miniprep Kit (50). Ambos métodos ofrecen rendimientos comparables, como se muestra en la figura « Manipulación de concentrados de soluciones tampón».

Ver figuras

QIAwave Plasmid Miniprep Kit performance

Spin column handling options: microcentrifuge.

Spin column handling options: vacuum manifolds.

Spin column handling options: automated system.

Complete digestion with various restriction enzymes.

Handling of buffer concentrates.

Principio

Las QIAprep 2.0 Spin Columns contienen una membrana de sílice única que une hasta 20 μg de ADN en presencia de una concentración alta de sal caotrópica y permite la elución en un volumen pequeño de solución tampón de baja salinidad. La tecnología de membrana de QIAprep elimina la extracción con fenol y cloroformo y la precipitación con alcohol que requieren de mucho tiempo, así como los problemas e inconvenientes relacionados con las resinas sueltas y los residuos pastosos. El ADN plasmídico ultrapuro eluido a partir de las QIAprep 2.0 Spin Columns está inmediatamente listo para su uso y no es necesario precipitarlo, concentrarlo o desalarlo.

Procedimiento

La purificación de plásmidos de ADN utilizando el QIAwave Plasmid Miniprep sigue un sencillo procedimiento de unión, lavado y elución (consulte el diagrama « Procedimiento QIAwave plasmid Miniprep»).

1. Lise los cultivos bacterianos y limpie los lisados mediante centrifugación.

2. Añada los lisados limpios a las QIAprep 2.0 Spin Columns. En este punto, la membrana de sílice absorbe el ADN plasmídico y se eliminan las impurezas.

3. El ADN puro se eluye en un volumen pequeño de solución tampón de elución o agua.

Además de usarse para la purificación de ADN plasmídico de E. coli, el QIAwave Plasmid Mini Kit se puede utilizar para purificar el ADN plasmídico de Saccharomyces cerevisiae, Bacillus subtilis y Agrobacterium tumefaciens. Póngase en contacto con nuestro equipo de servicio técnico o con su distribuidor local si necesita protocolos para estas aplicaciones.

Las soluciones tampón QIAwave se presentan como concentrados que pueden reconstituirse fácilmente añadiendo agua o etanol; consulte el manual de uso para obtener más información. Las QIAwave QIAprep 2.0 Spin Columns y los tubos de residuos vienen en bolsas individuales y deben montarse previamente antes de iniciar el protocolo. Esto lleva un poco más de tiempo, pero reduce los residuos de plástico.

El QIAwave Plasmid Miniprep Kit puede automatizarse en el QIAcube Connect utilizando los protocolos del QIAprep Spin Miniprep Kit.

Ver figuras

QIAwave Plasmid Miniprep procedure.

Aplicaciones

El QIAwave Plasmid Miniprep Kit aporta rendimiento reproducible de ADN ultrapuro apto para su uso en la mayoría de las aplicaciones, incluidas:

PCR
Digestión de restricción
Ligación y transformación
Secuenciación
Cribado

Datos y cifras de respaldo

QIAwave Plasmid Miniprep Kit (50) ACT environmental impact factor label US.

This ACT environmental impact factor label evaluates and scores the QIAwave Plasmid Miniprep Kit (50) for plasmid DNA isolation on several sustainability criteria. Scores are 1–10 except for energy and water consumption, which is scored as 1 point per kWh. or gallon, respectively. A low score means a lower environmental impact. The QIAwave Plasmid Miniprep Kit (50) has a 36.8% lower EIF in the US than the equivalent standard kit, the QIAprep Spin Miniprep Kit (50).

Specifications

Features	Specifications
Applications	Secuenciación fluorescente y radioactiva (incluida la secuenciación capilar), ligación, clonación, transformación, etc.
Processing	Manual (centrifugation or vacuum)
Plasmid type	High-copy, low-copy, cosmid DNA
Culture volume/starting material	1–10 mL culture volume
Elution volume	50 µL (minimal)
Technology	Silica 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 run	1–24 samples per run

Recursos

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.

More eco-friendly alternatives to our standard kits for extracting DNA and/or RNA

FAQ

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

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

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

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

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

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

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

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

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

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

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

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

Yes, please follow either of the User-Developed Protocols:

FAQ ID -896

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

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

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

Yes, please check the QIAprep Spin Miniprep Kit high-yield protocol for purification of up to 30 µg plasmid DNA.

FAQ ID -3102

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

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

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

Yes, please follow the User-Developed Protocol 'Isolation of plasmid DNA from Agrobacterium using the QIAprep Spin Miniprep Kit; spin procedure' (PR03s).

FAQ ID -898

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

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

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

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