The NGS DNA Fragmentation & Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality libraries for next generation sequencing. The kit uses intact genomic DNA (EDTA-free DNA or DNA resuspended in TE buffer) as input DNA without an additional DNA fragmentation step. Our technology provides a fast and simple workflow. DNA libraries can be generated around 2 hours with only 10 min hands-on time. Library multiplexing is possible.
NGS DNA Fragmentation & Library Prep Kit Workflow
The incorporation of DNA fragmentation in the kit makes it possible to directly use intact genomic DNA as input DNA without the need of mechanical DNA shearing or enzymatic DNA fragmentation. The NGS DNA Fragmentation & Library Prep Kit does not generate sequencing bias as compared to library using mechanical sheared DNA as input. Sequence coverage is also consistent between enzymatic shearing and mechanical shearing. The library size is inversely correlated with the incubation time of step 1 at 20°C.
Three index types are available for the kit of the illumina platform:
Non-index (Cat.# 30026): Libraries do not have index.
Index (Cat.# 30028): Each of the index primers contains a unique index sequence of 6 bases. Library multiplexing for 48 samples is possible. Index information can be downloaded here.
Unique dual index (Cat.# 30030): Library multiplexing for 96 samples is possible. With the unique features of our 4-Base Difference Index System, the index sequence is 8-base long and each index has 4 bases different from others. Our unique dual index primers effectively identify sequencing errors such as index hopping, mis-assignment of reads, and de-multiplexing errors etc. The unique dual index primers set consists of 96 pre-mixed unique pairs of index primers. Index information can be downloaded here.
Indexes are available for the MGI platform kits (Cat.# 34028).
Kit features:
Library conversion efficiency: 100 ng, 300 ng and 500 ng of intact genomic DNA were used as input.
The library size is inversely correlated with the incubation time of step 1 at 20°C.
NGS data comparison: enzymatic shearing versus mechanical shearing
Enzymatic shearing
• DNA shearing and library prep: BioDynami NGS DNA Fragmentation & Library Prep Kit
Mechanical shearing
• DNA shearing: Covaris sonication
• Library prep: BioDynami NGS DNA Library Prep Kit.
The NGS DNA Fragmentation & Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality libraries for next generation sequencing. The kit uses intact genomic DNA (EDTA-free DNA or DNA resuspended in TE buffer) as input DNA without an additional DNA fragmentation step. Our technology provides a fast and simple workflow. DNA libraries can be generated around 2 hours with only 10 min hands-on time. Library multiplexing is possible.
Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.
The series of nucleic acid columns produced by Magen Biotech are based on carefully selected imported glass fiber membranes (GF/B, GF/D, GF/F). Columns production processes such as polypropylene injection molding materials, injection molding process, and downstream membrane packing and compression rings are strictly controlled. This is to ensure that the column has extremely high adsorption capacity and long-term stability. Compared with conventional products on the market, Magen’s columns are with varieties, and binding rate will not change when stored at room temperature for 4 years.
Specifications
Features | Specifications |
Recommended application | Small amounts of nucleic acid isolation, viral nucleic acid from cell free samples |
Preservation conditions | Room temperature |
Stability | Up to 4 years |
Filter membrane | High quality glass fiber filter GF/F, 2 layers |
Membrane aperture | 0.7μm |
Maximum binding yield of plasmid | 30 μg |
Maximum yield of alcohol mediated Binding | 100 μg |
Single liquid carrying capacity of column | 900 μl |
Minimum elution volume | 80 μl |
Withstand centrifugal force | 5,000 x g |
Centrifuge | Low speed centrifuge, Swing out Rotor, can placed a height of 6.5cm square, (height of HiPure DNA Plate & 1.6ml Collection Plate: height, 6.2cm) |
Adsorption Mechanism
Based on the negatively charged DNA skeleton, it has a high affinity for positively charged glass fibers. In high salt and ethanol solutions, DNA/RNA binds to glass fiber and interacts with hydrophilic matrix on silica through hydrogen bond. DNA/RNA is tightly bound. All pollutants can be removed by washing solution. At high salt concentration, nucleic acids selectively bind to silica gel membrane, while other pollutants, mainly proteins, are removed by membrane washing.
Ordering information
CAT.No. | Product Name | Package |
C13130 | HiPure DNA Plate (2 x GF/F)with 1.6ml Collection Plate | 10/Bag |
Item No. | Product Name | Membrane type/number of layers | Collection tubes | Plasmid DNA binding capacity (Physical adsorption) | gDNA/RNA binding capacity (Alcohol-mediated adsorption) | Minimum Elution volume | Liquid volume capacity |
C13010 | HiPure DNA Nano Column | 2 layers GF/F | 2ml without cap | 5μg | 20μg | 10μl | 700μl |
C13011 | HiPure DNA Micro Column | 3 layers GF/F | 2ml without cap | 10μg | 50μg | 15μl | 700μl |
C13100 | HiPure DNA Mini Column I | 2 layers GF/B | 2ml without cap | 15μg | 100μg | 30μl | 700μl |
C13110 | HiPure DNA Mini Column II | 4 layers GF/B | 2ml without cap | 35μg | 200μg | 50μl | 800μl |
C13111 | HiPure RNA Mini Column | 3 layers GF/B | 2ml without cap | 30μg | 200μg | 30μl | 800μl |
C13112 | HiPure Viral Mini Column | 3 layers GF/F | 2ml without cap | 30μg | 200μg | 30μl | 800μl |
C13113 | HiPure CFDNA Mini Column | 3 layers GF/F,1 layer GF/B | 2ml without cap | 30μg | 200μg | 30μl | 800μl |
C13120 | HiPure DNA Midi Column | 4 layers GF/B | 15ml Centrifuge tube | 125μg | 1mg | 500μl | 4ml |
C13121 | HiPure DNA Midi Column III | 8 layers GF/B | 15ml Centrifuge tube | 250μg | 1mg | 500μl | 4ml |
C13122 | HiPure DNA Maxi Column | 4 layers GF/B | 50ml Centrifuge tube | 500μg | 5mg | 1000μl | 20ml |
C13123 | HiPure DNA Maxi Column III | 8 layers GF/B | 50ml Centrifuge tube | 1mg | 5mg | 1000μl | 20ml |
C13124 | HiPure DNA Maxi Column C | 8 layers GF/B | 50ml high speed Centrifuge tube | 1mg | 5mg | 700μl | 12ml |
C13130 | HiPure DNA Plate | 2 layers GF/F | 1.6ml Plate | 30μg | 100μg | 80μl | 900μl |
C13131 | HiPure gDNA Plate | 2 layers GF/B | 1.6ml Plate | 30μg | 100μg | 80μl | 900μl |
Note: GF/B pore size is for 1.0μM glass fiber membrane; GF/F pore size is for 0.7μm glass fiber membrane.
Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.
The HiPure Plasmid EF Mini Kit combine the power of HiPure technology with Magen’s innovative endotoxin removal technology to deliver high-quality plasmid DNA with low endotoxin levels foruse in eukaryotic transfection, and in vitro experiments. The HiPure plasmid endo-free system uses a specially formulated buffer that prevents endotoxin molecules from binding to the surface of the HiPure matrix. Endotoxin contamination lowers transfection efficiencies for endotoxin sensitive celllines. For gene therapy, endotoxin contamination should be of major concernsince endotoxins have the potential to cause fever, endotoxin shock syndrome, and interfere with in vitro transfection into immune cells.
Specifications
Features | Specifications |
Main Functions | Isolation up to 80µg endotoxin-free plasmid DNA from 5-15ml bacterial culture. Recommend for low copy vector, Thoroughly remove RNA |
Applications | Cell transfection, animal injection, etc. |
Purification method | Mini spin column |
Purification technology | Silica technology |
Process method | Manual (centrifugation or vacuum) |
Sample type | Low copy plasmid vector |
Sample amount | 5-15ml LB |
Yield | 10-70μg |
Elution volume | ≥75μl |
Time per run | ≤40 minutes |
Liquid carrying volume per column | 800μl |
Binding yield of column | 70μg |
The HiPure Plasmid procedure is based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt. The unique silica membrane used in the kit completely replaces glass or silica slurries for plasmid DNA minipreps. The procedure consists of 3 basic steps: Preparation and clearing of a bacterial lysate by alkaline method,then transfer the supernatant to column to bind DNA. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).
Advantages
Kit Contents
Contents | P115402 | P115403 |
Purification Times | 50 Preps | 250 Preps |
RNase A | 5 mg | 20 mg |
Buffer P1 | 30 ml | 140 ml |
Buffer P2 | 30 ml | 140 ml |
Buffer LEN3 | 15 ml | 70 ml |
Buffer LN4 | 50 ml | 250 ml |
Buffer LN5 | 30 ml | 140 ml |
Buffer PW1 | 30 ml | 140 ml |
Buffer PW2 | 12 ml | 50 ml |
Elution Buffer | 15 ml | 30 ml |
HiPure DNA Mini Columns III | 50 | 250 |
2 ml Collection Tubes | 50 | 250 |
Storage and Stability
The kit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve. After addition of RNase A,Buffer P1 is stable for 6 months when stored at 2–8°C.
For any technical problems or customized products, please contact us.
F&Q about Endotoxin-free Plasmid Extraction Kit — P1156 ←click here
The HiPure Plasmid EF Mini Kit combine the power of HiPure technology with Magen’s innovative endotoxin removal technology to deliver high-quality plasmid DNA with low endotoxin levels foruse in eukaryotic transfection, and in vitro experiments. The HiPure plasmid endo-free system uses a specially formulated buffer that prevents endotoxin molecules from binding to the surface of the HiPure matrix. Endotoxin contamination lowers transfection efficiencies for endotoxin sensitive celllines. For gene therapy, endotoxin contamination should be of major concernsince endotoxins have the potential to cause fever, endotoxin shock syndrome, and interfere with in vitro transfection into immune cells.
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