QIAwave Plasmid Miniprep Kit

For a more eco-friendly alternative to our standard kit for extracting up to 20 μg molecular biology grade plasmid DNA

Features

  • Plasmid DNA quality and performance identical to the QIAprep Spin Miniprep Kit
  • 15% less plastic and 15% less cardboard compared to the QIAprep Spin Miniprep Kit
  • Reusable Waste Tubes made from 100% post-consumer recycled plastic
  • Buffer concentrates that use 48–80% less plastic than our standard buffers
S_1177_0_QF_QIAwave

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QIAwave Plasmid Miniprep Kit (250)

Cat. No. / ID: 27206

QIAprep 2.0 Spin Columns, Waste Tubes (2ml), Reagents
€379.00
KitKit component
QIAwave Plasmid Miniprep Kit
Waste Tubes
The QIAwave Plasmid Miniprep Kit (250) is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Product Details

The QIAwave Plasmid Miniprep Kit is an eco-friendlier version of our standard QIAprep Spin Miniprep Kit. The QIAwave Kit uses 15% less plastic and 15% less cardboard than our standard kit and offers waste tubes made 100% post-consumer recycled plastic that you can reuse throughout the procedure. QIAwave buffers also come as concentrates, reducing the amount of plastic by 48–80% per bottle. To save paper, there are no printed protocols in the kit. Instead, you can download the protocols you need from the resources list or by simply scanning the QR code inside the kit box lid.

So, while the kit packaging and components of our QIAwave Kit may look different, it’s as easy to use as our standard kit, and the chemistry and performance are identical.

In partnership with My Green Lab, we've also assessed the environmental impact of this kit. My Green Lab ACT (accountability, consistency, and transparency) environmental impact factor labels are designed to evaluate and score products on several sustainability criteria. These include:

  • manufacturing
  • responsible chemical management
  • sustainable content within products and packaging materials
  • disposal of the packaging at the end of life

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 impact.

You can view the ACT labels for this kit in the Resources section below.

The QIAwave Plasmid Miniprep Kit is designed for isolation of up to 20 μg high-purity plasmid or cosmid DNA to use in routine molecular biology applications, including fluorescent and radioactive sequencing and cloning. You can also achieve even higher yields (up to 30 μg) using the High-Yield Supplementary Protocol. For optimal results, we recommend using this kit together with the QIAvac 24 Plus. And if you have too many samples to prep manually, you can automate your sample extraction on the QIAcube Connect.

Performance

The performance between our QIAwave Plasmid Miniprep Kit and QIAprep Spin Miniprep Kit is identical because the chemistry is the same. We’ve also shown that both kits outperform competitors’ kits (see figure " QIAwave Kit performance").

The QIAwave Plasmid Miniprep Kit allows you to purify up to 20 μg molecular biology grade plasmid DNA or cosmid DNA to use in routine molecular biology applications such as PCR, sequencing and cloning.

The QIAprep 2.0 Spin Columns are so versatile that you can use them in microcentrifuges, on vacuum manifolds or on the QIAcube Connect (see figures "QIAprep 2.0 Spin Column handling options  A,  D, and  E"). Th vacuum procedure makes handling simpler and sample processing faster. The QIAprep 2.0 Spin Columns can also be vacuum processed using the QIAvac 24 Plus or any other commercial manifold with luer connectors. You can also fully automate the extraction protocol using the QIAcube Connect.

Format Spin columns
Purification module QIAprep 2.0 Spin Columns
Throughput 1–24 samples
Preparation time 24 minipreps in 30 minutes
Equipment required Microcentrifuge or vacuum manifold; fully automatable using the QIAcube Connect
Lysate clearing Centrifugation
Capacity of column reservoir 800 µl
Minimum elution buffer volume 50 µl
Culture volume for high-copy plasmids 1–5 ml
Culture volume for low-copy plasmids/cosmids 1–10 ml

Purified DNA can be used in restriction digestion (see figure "Complete digestion with various restriction enzymes").

See figures

Principle

The QIAprep 2.0 Spin Columns contain a unique silica membrane that binds up to 20 μg DNA in the presence of a high concentration of chaotropic salt and allows elution in a small volume of low-salt buffer. QIAprep membrane technology eliminates time-consuming phenol-chloroform extraction and alcohol precipitation, as well as the problems and inconvenience associated with loose resins and slurries. High-purity plasmid DNA eluted from QIAprep 2.0 Spin Columns is immediately ready to use – there is no need to precipitate, concentrate, or desalt.

Procedure

DNA plasmid purification using the QIAwave Plasmid Miniprep follows a simple bind-wash-elute procedure (see flowchart " QIAwave plasmid Miniprep procedure").

1. Lyse bacterial cultures and clear the lysates using centrifugation.

2. Add the cleared lysates to the QIAprep 2.0 Spin Columns. At this point, the plasmid DNA adsorbs to the silica membrane and impurities are washed away.

3. The pure DNA is then eluted in a small volume of elution buffer or water.

In addition to plasmid purification from E. coli, the QIAwave Plasmid Mini Kit can be used to purify plasmid DNA from Saccharomyces cerevisiae, Bacillus subtilis, and Agrobacterium tumefaciens. Contact our Technical Services Team or your local distributor if you need protocols for these applications.

QIAwave buffers come as concentrates that you can easily reconstitute by adding water and/or ethanol; please check the handbook for details. QIAwave QIAprep 2.0 Spin Columns and Waste Tubes come in individual bags and need to be preassembled before you start the protocol. This takes a little extra time, but it does reduce plastic waste.

If you have too many samples to process manually, you can automate the purification process using the DNeasy mini spin prep columns on the QIAcube Connect.

See figures

Applications

The QIAwave Plasmid Miniprep Kit provides reproducible yields of high-purity DNA suitable for use in most applications, including:

  • PCR
  • Restriction digestion
  • Ligation and transformation
  • Sequencing
  • Screening

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsFluorescent and radioactive sequencing (including capillary sequencing), ligation, cloning, 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

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
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
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?
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
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
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 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