Cat. No. / ID: 27204
The QIAwave Plasmid Miniprep Kit is an eco-friendlier version of our standard QIAprep Spin Miniprep Kit. The QIAwave Kit uses up to 22% less plastic and up to 14% less cardboard than our standard kit and offers waste tubes made from 100% post-consumer recycled plastic that you can reuse throughout the procedure. QIAwave buffers come as concentrates, reducing the amount of plastic by up to 93% per bottle. To save paper, there are no printed protocols in the kit. You can download the protocols from the resources list or scan the QR code inside the kit box. Although 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.
Please be aware that you will need sterile glass bottles to store the reconstituted buffers.
In partnership with My Green Lab, we've assessed the environmental impact of this kit. My Green Lab ACT labels are designed to evaluate and score products on several sustainability criteria. These include:
• 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 (see figures "QIAwave Plasmid Miniprep Kit ACT environmental impact factor label US, EU and UK").
The QIAwave Plasmid Miniprep Kit is designed to isolate up to 20 μg high-purity plasmid or cosmid DNA for routine molecular biology applications, including fluorescent and radioactive sequencing and cloning. You can achieve even higher yields (up to 30 μg) using the High-Yield Supplementary Protocol. We recommend using this kit with the QIAvac 24 Plus for optimal results.
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.
|Purification module||QIAprep 2.0 Spin Columns|
|Preparation time||24 minipreps in 30 minutes|
|Equipment required||Microcentrifuge or vacuum manifold; fully automatable using the QIAcube Connect|
|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").
We have also compared plasmid DNA yields obtained using the QIAwave Plasmid Miniprep Kit (50) buffer, prepared by pouring or pipetting, and the QIAprep Spin Miniprep Kit (50) standard buffers. Both methods result in comparable yields as shown in the figure “ Handling of buffer concentrates”.
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.
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.
The QIAwave Plasmid Miniprep Kit can be automated on the QIAcube Connect using the QIAprep Spin Miniprep Kit protocols.
The QIAwave Plasmid Miniprep Kit provides reproducible yields of high-purity DNA suitable for use in most applications, including:
|Applications||Fluorescent and radioactive sequencing (including capillary sequencing), ligation, cloning, transformation 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)|
|Time per run or prep per run||<30 minutes|
|Samples per run (throughput)||1–24 samples per run|
|Number of preps per run||1–24 samples per run|
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.
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|
|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|
* 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.
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.
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.
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.
Yes, please follow either of the User-Developed Protocols:
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.
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.
Yes, please follow the User-Developed Protocol 'Isolation of plasmid DNA from Agrobacterium using the QIAprep Spin Miniprep Kit; spin procedure' (PR03s).
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.
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.
The components of the SOC medium are:
*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.