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Macherey & Nagel GmbH, Germany

Announcing the launch of Nucleic Acid Purification Kits.
DNA / RNA Isolation & Purification Kits

Ultra-fast purification of genomic DNA from blood

DNA-free total RNA Purification in the 96-well format

MACHEREY-NAGEL has developed the NucleoSpinMulti-96 RNA kit for the rapid parallel purification of highly pure total RNA from 10 - 5 x 105 cultured cells in the 96-well format. Based on silica membrane technology, 2 x 96 samples can be processed in about 1 hour. Removal of residual DNA is achieved by incubation with DNase I (included in the kit), which is directly applied onto the silica membrane during the preparation. Elution of total RNA with the small elution volume of 25 – 50 µl leads to high RNA concentrations and thus to high sensitivities. Optimised chemistry and plate design prevent cross-contamination and lead to reproducible yields of up to 5 µg of high quality total RNA which is free of degradation and ready-to-use for multiple RT-PCR´s. The kit can be adapted to common pipetting robots.

Endotoxin-free plasmid DNA - small and large scale purification with NucleoBond Kits from MACHEREY-NAGEL

MACHEREY-NAGEL's new NucleoBond PC 500 EF Kit is designed for purification of ultra-pure endotoxin-free plasmid DNA. The kit allows the isolation of up to 500 µg DNA from about 50 – 100 ml E. coli culture. Based on anion exchange chromatography, the working procedure requires no additional step and allows an easy handling with a minimum hands-on-time. The purified plasmid DNA with less than 0.05 EU/µg DNA is suitable for transfection of sensitive cell lines and gene therapy. Preparative isolation of plasmid DNA is now possible with the NucleoBond PC Prep 100 Kit, yielding up to 100 mg of plasmid DNA from 5-20 litres of E. coli culture. Resulting plasmid DNA has an A260/280-ratio of 1.80-1.90, shows a structural integrity of at least 90% supercoiled DNA and has an endotoxin level of less than 0.1 EU/µg DNA. Therefore, the ultrapure plasmid DNA prepared with NucleoBond PC Prep 100 Kit is suitable for any sensitive downstream application, like transfection, gene therapy and vaccination. NucleoBond AX columns contain a silica-based, outstanding anion exchanger (covered by European patent E.P. 0496822), developed and manufactured by MACHEREY-NAGEL.

Highly sensitive purification of viral RNA / DNA

With the NucleoSpin RNA Virus kit, MACHEREY-NAGEL has developed a rapid and easy procedure which is suitable for the extraction of viral nucleic acids from 150 µl serum, plasma or other cell-free biological fluids. The NucleoSpin RNA Virus kit features a special type of silica membrane which has a high binding capacity for RNA / DNA, especially for viral RNA. The recovery rate of viral RNA is >90% and even highly diluted samples containing theoretically about 30 – 60 RNA copies/ml lead to positive signals with nested PCR detection. Processing of larger samples, e.g. pooled samples, is now possible with the NucleoSpin RNA Virus F kit of MACHEREY-NAGEL. The specially designed funnel column allows due to the closed system a cross-contamination free handling of up to 1 ml serum or plasma within one loading step. Because of the small diameter of the silica-membrane the elution volume is still between 50 – 100 µl resulting in a highly concentrated sample ready-to-use for any amplification process like RT-PCR. For the isolation of viral DNA with highest sensitivities we recommend the NucleoSpin Blood kit (e.g. detection of about 100 copies/ml of HBV DNA with nested PCR).

Medium and High-throughput purification of genomic DNA from Blood

MACHEREY-NAGEL has developed silica-membrane based kits for rapid parallel purification of genomic DNA from blood, serum, plasma and body fluids. The NucleoSpin Multi-8 Blood kit offers a 8-well strip system for a flexible handling of multiples of 8 samples. The convenient processing of multiples of 96 samples is achieved by using NucleoSpin Multi-96 Blood. Both kits allow purification of 4 – 12 µg genomic DNA from up to 200 µl samples in less than 30 minutes. With the NucleoSpinBlood method, genomic DNA is prepared from fresh or frozen human or animal blood samples treated with anticoagulants like citrate, EDTA or heparin. After efficient lysis contaminations are removed by washing with two different buffers. Highly pure genomic DNA is eluted under low ionic strength conditions with a final concentration of 40 – 120 ng/µl. The DNA is ready-to-use for enzymatic reactions and southern blotting. NucleoSpinMulti-8 and Multi-96 Blood kits are used in combination with a benchtop centrifuge like the NucleoSwing with deep buckets allowing processing of NucleoSpinMulti-96 Blood plates stacked on 96-well blocks.

NucleoBond PC Prep 100 Kit for the isolation of up to 100 mg of plasmid DNA

MACHEREY-NAGEL's new NucleoBond PC Prep 100 kit for large-scale plasmid purification yields up to 100 mg of plasmid DNA from 5 - 20 litres of E. coli culture. Resulting plasmid DNA with an A260/280-ratio of 1.80 - 1.90, shows a structural integrity of at least 90% supercoiled (ccc) DNA. Due to an endotoxin level of less than 0.1 EU/µg DNA, the ultrapure plasmid DNA prepared with NucleoBond PC Prep 100 kit is suitable for any downstream application like transfection and gene therapy. Harvested bacteria are lysed by an optimised alkaline lysis procedure. The crude lysate is cleared by filtration and applied to prepacked NucleoBond AX Prep 100 column by a peristaltic pump. After two washing steps, ultrapure plasmid DNA is eluted with a high-salt buffer. Following the precipitation with isopropanol, plasmid DNA is resuspended in Tris buffer (or water). NucleoBond AX columns contain a silica-based, outstanding anion exchanger developed and manufactured by MACHEREY-NAGEL.

NucleoSpin Plus for the preparation of up to 40 µg plasmid DNA

MACHEREY-NAGEL's improved NucleoSpin Plasmid Mini kit now yields up to 40 g plasmid DNA, more than any other spin column based kit. Up to 8 ml ON culture can be processed. As a manufacturer of silica and glass fiber filters MACHEREY-NAGEL was able to develop an optimized membrane for the isolation of a much higher amount of nucleic acids. The purification is based on the silica membrane-spin-technology. After alkaline lysis and precipitation of chromosomal DNA the plasmid DNA is isolated by the silica membrane in presence of a chaotropic salt. Contaminants are removed by specific washing steps. Plasmid DNA is eluted by a low-salt buffer and is ready-to-use for investigations like restriction analysis, cloning, hybridization, PCR and sequencing (AFS or conventional techniques).

NucleoSpin Virus L - a kit for the extraction of viral nucleic acids from up to 2 ml serum or plasma

MACHEREY-NAGEL has designed a new kit for the ultra-fast small-scale purification of genomic DNA from whole blood, serum, plasma and body fluids. The NucleoSpin Blood QuickPure kit is based on improved silica membrane technology. The speeding up of the procedure is achieved by only ONE combined washing/drying step instead of three working steps according to standard procedure. The hands-on-time is less than 10 minutes. Blood samples with any type of anticoagulant frozen or clotted can also be processed conveniently. From sample sizes of up to 200 µl the NucleoSpin Blood QuickPure kit allows typical yields of 4 – 6 µg highly pure genomic DNA. The DNA with an A260/280-ratio between 1.60-1.90 is ready-to-use for also sensitive (PCR) enzymatic reactions. Due to the small elution volume of 25 – 50 µl a high concentration of 40 – 120 ng DNA per µl can be achieved.

All available spin column devices for the purification of viral nucleic acids are able to handle 200 µl of serum or plasma in one loading step. Sometimes, especially if several samples have been pooled, sensitivity is too low for a subsequent amplification reaction. The only possibility is to use a larger volume of sample and to load the sample successively onto the spin column. This time consuming process is not user friendly because of the high potential of cross contaminations

For this reason MACHEREY-NAGEL has developed a new column for the processing of larger volumes. The so called "funnel column" of our NucleoSpin Virus L kit (L stands for large volume) allows the handling of up to 1 ml serum or plasma within one loading step. Due to the small diameter of the silica membrane, the elution volume is still between 50 -100 µl low salt buffer or water. This means a high concentrated sample can be obtained, directly suitable for any amplification process. Quantification analysis shows that even 20 copies HCV/ml can be detected

The funnel column can be centrifuged in a 50 ml polyethylene tube (included in the kit) which is closed separately. This avoids cross contaminations. The chemistry of the procedure is the same as in our NucleoSpin Virus kit. A guanidine isothiocyanate based buffer allows the lysis of the virus particles as well as the purification of the nucleic acids by a specially modified silica membrane. Two different elution buffers are used for the extraction of viral RNA and DNA molecules. Elution tubes (0.5 ml, included in the kit) are fitted directly to the outlet of the funnel column

8-12 samples (depending on the rotor) can be processed within less than 30 minutes. This allows to reduce significantly the bottleneck of processing larger volumes

NucleoBond PC Plasmid kits now include folded filters

MACHEREY-NAGEL is the manufacturer of the well-known NucleoBond AX kits for the purification of DNA and RNA (anion exchange chromatography). Now, all NucleoBond PC 100 (Midi), PC 500 (Maxi) and PC 2000 (Mega) Plasmid kits include the required number of folded filters without any additional costs. The use of folded filters allows the clarification of the bacterial lysats instead of a time consuming centrifugation. In just 10 minutes lysates can be filtered, leaving behind salt precipitates and cellular debris. The filtration step increases your plasmid integrity by minimizing possible DNA shearing that can occur during centrifugation or pouring lysates through syringe filters. The clarified lysate can be loaded directly onto a preequilibrated NucleoBond AX column.

NucleoSpin Multi 8 Plasmid and NucleoSpin Multi 8 Plus Plasmid kits for the purification of plasmid DNA

The NucleoSpin Multi 8 and the NucleoSpin Multi 8 plus Plasmid kit of MACHEREY-NAGEL allow the manual high throughput purification of up to 15 g plasmid DNA. These kits offer convenient processing of up to 96 samples in a highly flexible 8well format and in less than 60 minutes by the use of a special vacuum manifold (NucleoVac). The NucleoSpin Multi 8 plus Plasmid kit additionally contains 8well strips for the filtration of the bacterial lysates instead of the time consuming centrifugation.

Plasmid DNA is purified by a specially modified silica membrane in the presence of a chaotropic salt. This process is independent from the copy number and size of the plasmid DNA. Contaminants are washed away and are removed prior to elution of DNA. The plasmids are eluted into deep-well collection tubes or into a microtiter plate. In contrast to other procedures there is no need for resuspension of a silica matrix. Time consuming phenol/chloroform extractions or alcohol precipitations are also eliminated

Additionally to the 12 8well strips and the corresponding buffer solutions, each kit contains a one side sealed microtiter washing plate. This plate is placed into the vacuum manifold in order to prevent cross contaminations

Purified nucleic acids are ready to use for restriction analysis, cloning, hybridization and for sequencing (AFS or conventional techniques).

NucleoSpin Multi 8 Extract kit for the direct purification of PCR products

After the successful introduction of our 8well strips for the preparation of plasmid DNA, MACHEREY-NAGEL has developed this kit for the manual high throughput purification of up to 15 g PCR products. The kit offers convenient processing of up to 96 samples in a highly flexible 8well format and in less than 60 minutes by the use of a special vacuum manifold (NucleoVac).

Within each column a specially modified silica membrane allows the purification of PCR fragments ( 100 bp) in the presence of a chaotropic salt. Contaminants like primers and primer/dimers are washed away and are removed prior to elution of the PCR products. The PCR fragments are eluted into deep-well collection tubes or into a microtiter plate. In contrast to other procedures there is no need for resuspension of a silica matrix or for removal of mineral oil. Time consuming phenol/chloroform extractions or alcohol precipitations are also eliminated.

Additionally to the 12 8well strips and the corresponding buffer solutions, each kit contains a one side sealed microtiter washing plate. This plate is placed into the vacuum manifold in order to prevent cross contaminations.

NucleoSpin Multi 8 Extract can be used to purify PCR products over a broad range (100 bp - 10 kbp). The recovery rate of DNA is 75-90 %. Purified nucleic acids are ready-to-use for restriction analysis, cloning, hybridization and for sequencing (AFS or conventional techniques).

NucleoTrap mRNA for the preparation of 40 µg poly A+ mRNA

MACHEREY-NAGEL's new NucleoTrap mRNA kit is recommended for the isolation of up to 40 µg poly A+ mRNA starting from total RNA. The mRNA is not degraded and free of DNA contamination. The purification process is based on affinity chromatography: poly A+ mRNA is hybridized to oligo(dT) latex beads. The complex of mRNA-latex beads is washed and transfered to the NucleoSpin microfilter. After additional washing the mRNA is finally eluted with RNase-free water, and is ready-to-use for any downstream application like RT-PCR, construction of cDNA libraries, in vitro translation or RNase protection assays. The preparation of ultrapure mRNA is convenient and needs less than 30 minutes by the combination of latex beads, NucleoSpin microfilter and centrifugation steps.

NucleoSpin RNA II for the preparation of 20 µg DNA-free total RNA

MACHEREY-NAGEL's improved NucleoSpin RNA Mini kit yields up to 20 µg of DNA-free total RNA from cultured cells, tissue, bacteria, yeast and biological fluids. DNase I treatment eliminates genomic DNA contamination. The purification based on the silica membrane-spin-technology needs less than 30 minutes, and replaces problematic RNA isolation procedures. The lysis buffer containing GITC inactivates RNases, and creates appropriate RNA binding conditions. Contaminating DNA is removed by DNase I directly applied onto the silica membrane and supplied with the kit. DNase I and all contaminants are effectively removed by specific washing steps. Ultrapure total RNA is finally eluted with RNase-free water, and is ready-to-use for any downstream application like RT-PCR, Northern/dot/slot blotting, primer extension, array technology or RNase protection assays.

NucleoSwing Bench Top Centrifuge for high-throughput preparation of nucleic acids

NucleoSwing Z 383 Bench Top Bio-Centrifuge was developed by the cooperation of MACHEREY-NAGEL and HERMLE. The centrifuge is suitable for simultaneous 96-well format extraction of nucleic acids from biological samples. Thereby, 2 96-well plates of the NucleoSpin Multi 96 kits can be used in combination with deep-well blocks or MTP´s. This makes a fast high-throughput preparation possible, like the isolation of ultrapure genomic DNA (NucleoSpin Multi 96 Blood kit for whole blood, serum, plasma, biological fluids ; NucleoSpin Multi 96 Plant kit for plants, food, funghi ; NucleoSpin Multi 96 C+T kit for e.g. cells, tissue, bacteria, mouse tails) and the clean up of PCR products (NucleoSpin Multi 96 Extract kit). NucleoSwing Z 383 is a high-speed bench top centrifuge of up to 6.000 rpm and maximum RCF value of 6.000 x g. The low noise level of 72 dB(A) is achieved by the integrated rotor wind shield, making it possible to operate the centrifuge in any laboratory.