Introduction

The HiPure Plasmid EF Maxi Kit combines the power of HiPure technology with Magen’s innovative endotoxin removal technology (ETR) to deliver high-quality plasmid DNA with low endotoxin levels for use in eukaryotic transfection, and in vitro experiments. Up to 1000 µg high copy number plasmid DNA or 200 µg low copy number plasmid DNA can be purified from 150 mL overnight culture.

F&Q about Endotoxin-free Plasmid Extraction Kit — P1156 ←click here

Details

Specifications

Principle

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

• High purity – purified plasmid can be directly used in sequencing, enzyme digestion and PCR, etc.
• Fast – silica gel column purification is much faster than ion exchange
• High yield – up to 1.5mg plasmid can be binded in one column
• Ultra low endotoxin – content of endotoxin is less than 0.1 EU/μg, which can be directly used for cell transfection and animal injection, etc.

Kit Contents

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.

Purchase Guide

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 Maxi Kit combines the power of HiPure technology with Magen’s innovative endotoxin removal technology (ETR) to deliver high-quality plasmid DNA with low endotoxin levels for use in eukaryotic transfection, and in vitro experiments. Up to 1000 µg high copy number plasmid DNA or 200 µg low copy number plasmid DNA can be purified from 150 mL overnight culture.

Bis-Sulfone-PEG9-DBCO is a bis-alkylating labeling reagent that is selective for the cysteine sulfur atoms from a native disulfide. These reagents undergo bis-alkylation to conjugate both thiols derived from the two cysteine residues of a reduced native disulfide bond such as the interchain disulfide bonds of an antibody. The reaction results in covalent rebridging of the disulfide bond via a three carbon bridge leaving the protein structurally intact. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Bis-Sulfone-PEG9-DBCO is a bis-alkylating labeling reagent that is selective for the cysteine sulfur atoms from a native disulfide. These reagents undergo bis-alkylation to conjugate both thiols derived from the two cysteine residues of a reduced native disulfide bond such as the interchain disulfide bonds of an antibody. The reaction results in covalent rebridging of the disulfide bond via a three carbon bridge leaving the protein structurally intact. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Overview

• Preservation and isolation of both cf-DNA and cf-RNA from a single tube
• Fixative-free preservative, no cross-linking of DNA
• Preserve cf-DNA/ct-DNA for 30 days at ambient temperature and for up to 8 days at 37°C
• Preserve cf-RNA for 30 days at ambient temperature
• Preserve Circulating Tumour Cells (CTCs) for 14 days at ambient temperature
• No plasma volume loss after shipping/transportation
• Prevent hemolysis allowing better separation of plasma
• Prevent apoptosis of blood cells and fragmentation of genomic DNA
• Produce high quality/quantity of plasma cf-DNA/ct-DNA/cf-RNA
• Vacuumed to draw 8.7 mL of blood in 10 mL tubes
• The procedure for plasma isolation using this kit is compliant with ISO 20186-3:2019

Norgen’s cf-DNA/cf-RNA Preservative Tubes are closed, evacuated plastic tubes for the collection and the preservation of cf-DNA, circulating tumor DNA, cf-RNA and circulating tumor cells in human whole blood samples during storage and shipping. Using these tubes in combination with Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Kits or Norgen’s Plasma/Serum RNA Purification Kits enables the purification and concentration of an inhibitor-free cf-DNA/ct-DNA/cf-RNA in a very small elution volume. Plasma recovered from Norgen’s cf-DNA/cf-RNA Preservative Tubes is also compatible with any other cf-DNA and/or cf-RNA purification method. The purified cf-DNA, ct-DNA and cf-RNA are compatible with any downstream application assays, including PCR, qPCR, rt-qPCR, methylation-sensitive PCR, Southern Blot analysis, gene expression analysis, microarrays and NGS.

This product is for research use only and not for use in diagnostic procedures.

Performance

Norgen’s cf-DNA/cf-RNA Preservative Tubes abridge the collection/processing of whole blood for the subsequent purification of cf-DNA and/or cf-RNA. The cf-DNA/cf-RNA Preservative Tubes preserve and inhibit the programmed cell-death (apoptosis) of the blood, hence preventing the release of intracellular DNA/RNA into plasma. Whole blood samples can be shipped and stored at room temperature for the subsequent processing of plasma. The cf/ct DNA and cf-RNA levels are stable for up to 30 days at room temperature. The cf-DNA is also stable for up to 8 days at 37°C. When blood collected on Norgen’s cf-DNA/cf-RNA tubes is processed for plasma recovery no buffy coat will be generated and the Circulating tumor cells (CTCs) will be located at the bottom of the tube with the blood sediment. CTCs are stable for 14 days at room temperature where it can be extracted from the blood sediment or from whole preserved blood for cell sorting or for cf/DNA or cf-RNA purification.

Plasma samples produced from the whole blood sample collected into Norgen’s cf-DNA/cf-RNA Preservative Tubes and purified using Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Kits or Norgen’s Plasma/Serum RNA Purification Kits yield concentrated, high-quality and inhibitor-free cf-DNA/ct-DNA or cf-RNA for any downstream application.

Workflow

NOTE: This product is for research use only. For the CE marked version, see Cat. Dx63950

Details

Supporting Data

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Storage Conditions and Product Stability

1. When stored at 15-30°C, unfilled Norgen cf-DNA/cf-RNA Preservative Tubes are stable through the expiration date.  See tube label for expiry date.
2. Short-term storage/shipping at -20°C to 37°C for up to 10 days is acceptable for unfilled Norgen cf-DNA/cf-RNA Preservative Tubes.
3. Do not freeze unfilled Norgen cf-DNA/cf-RNA Preservative Tubes for long term storage over 10 days. Freezing unfilled tubes may lead to loss of vacuum and precipitation may occur.
4. Cell-free DNA in blood samples collected in Norgen’s cf-DNA/cf-RNA Preservative Tubes is stable for up to 30 days when stored at room temperature (15-25°C)
5. Cell-free DNA in blood samples collected in Norgen’s cf-DNA/cf-RNA Preservative Tubes is stable for up to 8 days when stored at 37ºC.
6. Cell-free RNA in blood samples collected in Norgen’s cf-DNA/cf-RNA Preservative Tubes is stable for up to 30 days when stored at room temperature (15-25°C)
7. Circulating tumor cells (CTCs) in blood samples collected in Norgen’s cf-DNA/cf-RNA Preservative Tubes are stable for up to 14 days at room temperature (15-25ºC).