Propargyl-PEG3-NHS ester is a Click Chemistry reagent with a propargyl group and an NHS ester group. The propargyl group can react with biomolecules containing azide group via copper catalyzed Click Chemistry reaction. The NHS ester is an amine reactive group which can be used for derivatizing peptides, antibodies, amine coated surfaces etc. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Propargyl-PEG3-NHS ester is a Click Chemistry reagent with a propargyl group and an NHS ester group. The propargyl group can react with biomolecules containing azide group via copper catalyzed Click Chemistry reaction. The NHS ester is an amine reactive group which can be used for derivatizing peptides, antibodies, amine coated surfaces etc. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
m-PEG4-DBCO is a click chemistry reagents which can enable so called copper-free Click Chemistry with azide to form a stable triazole. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
m-PEG4-DBCO is a click chemistry reagents which can enable so called copper-free Click Chemistry with azide to form a stable triazole. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
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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Library size selection is an enrichment of a specific range of library sizes for NGS library preparations. The NGS library preparation is related to the quality of the sequencing data. Precise NGS library size selection can increase sequencing efficiency, improve data quality, and reduce costs.
There are two types of sequencing technologies: short-read sequencing and long-read sequencing. Short-read sequencing uses DNA libraries that contain small insert DNA fragments of similar sizes, usually several hundred base pairs. The sequencing efficiency can be improved if the DNA size selection is in the right range. Cat.# 20104S and 20104L are the best kits for NGS library size selection of illumina paired-end 100 (PE100) sequencing with 100-200 bp library inserts; Cat.# 20105S and 20105L are the best kits for NGS library size selection of illumina paired-end 150 (PE150) sequencing with 150-300 bp library inserts; and Cat.# 20106S and 20106L are the best kits for NGS library size selection of illumina paired-end 300 (PE300) sequencing with 300-600 bp library inserts.
Long-read sequencing uses a large DNA fragment as input and makes very long reads. Usually, library size selection is preferred to remove smaller fragments. Cat.# 20110S and 20110L are the best kits for long-read sequencing size selection with DNA sizes >5 kb, and Cat.# 20111S and 20111L are the best kits for long-read sequencing size selection with DNA sizes >10 kb.
The magnetic beads, or SPRI (Solid Phase Reversible Immobilization) beads, is well used for the purification of DNA due to their reversible DNA binding. The NGS library can be size-selected by the magnetic beads or SPRI beads. The properties of the magnetic beads can be changed for a specific range of DNA binding. The contaminants and other unwanted components in the libraries can also be removed during size selection.
Specific ranges of NGS libraries can be selected using magnetic beads with different buffer compositions. The first DNA-beads binding step, also called the right-side clean-up, removes large NGS library fragments. The large NGS library fragments that bind to the beads are discarded with the beads pellet. The desired NGS library fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-DNA binding, also called the left-side clean-up. After the rinsing step, the NGS library fragments with the dual selection are eluted in water or an appropriate buffer. The magnetic beads method has great advantages over time-consuming column purification and tedious gel-based purification.
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Library size selection for long-read sequencing only requires a single clean-up. In this case, only the large library fragments are bound to the beads, while other small library fragments are discarded with the supernatant. The selected larger library fragments are eluted in water or an appropriate buffer after the rinsing step.
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