Miniprep kit manual

We recommend QuickLyse Miniprep Kit Add to cart. The QuickLyse Miniprep Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease. Open lightbox. Bacterial cells are resuspended and lysed and DNA is bound in a single buffer, enabling fast DNA purification with minimal hands-on time. After a 3-minute lysis step, the clear lysate is applied directly to a QuickLyse Spin Column. Plasmid DNA binds to the column membrane and is eluted after a single wash step see flowchart " QuickLyse procedure ".

The isolated DNA can be used directly in standard applications, including automated sequencing, PCR, restriction analysis, and cloning. You are not authorized to download the resource. Sort options Sort alphabetically. Can the bacterial pellet in the QuickLyse Miniprep procedure be resuspended by pipetting up and down or shortening the vortexing time? FAQ ID What volume of bacterial culture can be processed using the QuickLyse Miniprep Kit?

What is the largest plasmid size that can be purified using the QuickLyse Miniprep Kit? Kit Handbooks. For purification of sequencing grade plasmid DNA. Safety Data Sheets. Global contacts. 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. 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. The protocol is called: 'Purification of plasmid DNA prepared by other methods'. 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. 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 composition of Buffer P1 is:. Sterilize the final solution by passing it through a 0.

Ensure that isopropanol is used at room temperature for precipitation. Some bacterial strains, such as TG1 and JM, naturally produce a high level of carbohydrates.

However, carbohydrate contamination may also be observed when using other strains. The most common cause of this problem is over-growth of bacterial cultures. Clumps that occur after addition of Buffer P2 in a bacterial lysate containing LyseBlue reagent indicate poor resuspension of the bacterial cell pellet in Buffer P1. This handling error leads to inefficient cell lysis, and incomplete precipitation of SDS, cell debris, and genomic DNA. When resuspending the cell pellet, vortexing longer or resuspending the pellet by pipetting up and down can help.

To overcome this, continue mixing the solution by inverting it gently until a homogeneous blue suspension is achieved. Reorder now! Reorder from your past orders in just one click. Order by Quote. Quote Number. Add quote number from your quote document. Customer Number. Add customer number from your quote document. To remove a quote go to the Cart. View Quote Example. Catalog Number. Looking for a quick way to design experiments? Try the Workflow Configurator. A convenient tool to build experimental workflows and find products to match your needs.

Log Out. Show More. For high-purity plasmid minipreps: QIAprep 2. Log in to see your account pricing. Kit Column. This product is not intended for the diagnosis, prevention, or treatment of a disease. Spin column handling options — A. 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.

To enable faster and more convenient sample processing and analysis, gel loading dye is provided in the kit. GelPilot Loading Dye contains 3 tracking dyes xylene cyanol, bromophenol blue, and orange G to facilitate the optimization of agarose gel run time and prevent smaller DNA fragments migrating too far see figure " GelPilot Loading Dye ".

First, bacterial cultures are lysed and the lysates are cleared by centrifugation. The cleared lysates are then applied to the QIAprep 2. Impurities are washed away and pure DNA is eluted in a small volume of elution buffer or water.

The QIAprep procedure. Supporting data and figures The QIAprep procedure. Complete digestion with various restriction enzymes. Spin column handling options — C. Resources User-Developed Protocols 4. PDF 94KB. PDF 85KB. PDF 93KB. The procedure has been used successfully for isolation of a single-copy, PDF KB. Kit Handbooks 2. PDF 2MB. PDF 41KB. Quick-Start Protocols 2. PDF 51KB. PDF 48KB. Application Notes 2. Safety Data Sheets 1. Safety Data Sheets EN. Supplementary Protocols 1.

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Structure of the Escherichia coli OH7 heme oxygenase ChuS in complex with heme and enzymatic inactivation by mutation of the heme coordinating residue His The composition of buffer N3 is confidential. However, buffer N3 can be purchased separately. The catalogue number is How do I prepare Buffer MP?