Permagen’s SBS/ANSI footprint 24-Tube thermal block allows the use of up to twenty four 1.5 mL -2 mL Microcentrifuge tubes, or screw-cap vials for keeping sample temperatures consistent. Can be used for heating as well
SKU: CB024
Permagen’s SBS/ANSI footprint 24-Tube thermal block allows the use of up to twenty four 1.5 mL -2 mL Microcentrifuge tubes, or screw-cap vials for keeping sample temperatures consistent. Can be used for heating as well
These kits are designed for the rapid preparation of plasmid DNA from Escherichia coli.
Plasmid MiniPrep Kit
This kit is designed for the rapid preparation of plasmid DNA from small cultures of Escherichia coli using convenient spin columns. The plasmid DNA is preferentially purified from other cellular components such as genomic DNA and RNA. This kit is able to purify plasmids up to 13,000 bp in size, and the typical purification yield is up to 20 μg from 1.5 mL of bacterial culture. Purified DNA is of excellent quality for transformation, restriction enzyme digestion, sequencing and more. Also available in a 96-well format.
Plasmid MiniPrep Kit (Magnetic Bead System and High Throughput Magnetic Bead System)
Norgen’s Plasmid MiniPrep Kit (Magnetic Bead System) is designed for the rapid preparation of plasmid DNA from small batch cultures of Escherichia coli. Norgen’s magnetic beads bind DNA under optimized salt concentrations and release the bound DNA under low salt and slightly alkali conditions. The plasmid DNA is preferentially purified from other cellular components such as genomic DNA and RNA. The purified plasmids are fully digestible with all restriction enzymes tested, and are completely compatible with real-time PCR and NGS.
Norgen’s Plasmid MiniPrep Kit (Magnetic Bead System) is also available in a 96-well (HT) format for high throughput applications. Purification with the 96-well plates can be integrated with a robotic automation system.
Plasmid MaxiPrep Kit
This kit is designed for the rapid spin column preparation of plasmid DNA from up to 100 mL of Escherichia coli cultures. The kit allows for the isolation of plasmid DNA with final endotoxin levels of 0.1 EU/µg of DNA or less. The kit is able to purify plasmids up to 13,000 bp in size, and typical yields from a 100 mL culture for a high copy number plasmid are between 0.4 and 1.0 mg. The purified DNA is fully digestible with all restriction enzymes tested, and is completely compatible with manual or automated sequencing to achieve 95-100% accuracy.
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| Kit Specifications | |
| Column Binding Capacity | 25 µg |
| Size of Plasmids Purified | Up to 13,000 bp |
| Average Yield from 1.5 mL of Culture | Up to 20 µg |
| Time to Complete 10 Purifications | 30 minutes |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the of shipment. The RNase should be stored at -20°C upon arrival. The Resuspension Solution AZ should be stored at 4oC upon addition of RNase enzyme.
| Component | Cat. 13300 (50 preps) | Cat. 46400 (250 preps) | Cat. 46500 (4 preps) | Cat. 46600 (20 preps) | Cat. 60300 (50 preps) | Cat. 63000 (192 preps) |
|---|---|---|---|---|---|---|
| Resuspension Solution AZ | 12 mL | 60 mL | 20 mL | 100 mL | 12 mL | 2 x 20 mL |
| Lysis Buffer N | 40 mL | 80 mL | 40 mL | 2 x 80 mL | 40 mL | 2 x 40 mL |
| Buffer TN | 20 mL | 130 mL | 55 mL | 2 x 130 mL | 20 mL | 1 x 55 mL 1 x 20 mL |
| Wash Solution E | 12 mL | 2 x 18 mL | – | – | – | – |
| Elution Buffer K | 8 mL | 30 mL | – | – | 8 mL | 2 x 8 mL |
| Wash Solution J | – | – | 25 mL | 3 x 25 mL | – | – |
| Elution Buffer J | – | – | 24 mL | 120 mL | – | – |
| RNase A | 1 vial | 1 vial | 1 vial | 1 vial | 1 vial | 1 vial |
| Magnetic Bead Suspension | – | – | – | – | 1 x 1.1 mL | 4 x 1.1 mL |
| Spin Columns | 50 | 250 | – | – | – | – |
| Collection Tubes | 50 | 250 | – | – | – | – |
| DNA Maxi Spin Columns with Collection Tubes (Clear ring in column) | – | – | 4 | 20 | – | – |
| Maxi Spin Filter Columns with Collection Tubes (Grey ring in column) | – | – | 4 | 20 | – | – |
| 96-Well Plate | – | – | – | – | – | 2 |
| Elution Tubes (1.7 mL) | 50 | 250 | – | – | 50 | – |
| Elution Tubes (50 mL) | – | – | 4 | 20 | ||
| 96-Well Elution Plate | – | – | – | – | – | 2 |
| Adhesive Tape | – | – | – | – | – | 2 |
| Product Insert | 1 | 1 | 1 | 1 | 1 | 1 |
These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating and exosomal RNA using a convenient spin column method. These kits can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used, as heparin can significantly interfere with many downstream applications such as RT-PCR. The purified plasma/serum RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS. The EXTRAClean columns undergo stringent processing and rigorous quality control measures to minimize contamination traces, ensuring optimal results for sensitive applications such as NGS.
Cell-free circulating RNA, including exosomal RNA in plasma or serum, has the potential to provide biomarkers for certain cancers and disease states, and includes tumor-specific extracellular RNA in the blood. Exosomes are 40 – 100 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. The exosomes contain cell-specific proteins, lipids and RNAs, which are transported to other cells, where they can alter function and/or physiology. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. Recent work has demonstrated the presence of distinct subsets of microRNAs within exosomes which depend upon the tumour cell type from which they are secreted. For this reason, exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses, they can be efficiently recovered from biological fluids, such as plasma or serum.
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 50 μL to 200 μL. The purified plasma/serum RNA is eluted in a flexible final volume of 10 μL to 25 μL.
This utilizes a two-column method, and can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 250 μL to 1.5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 2 mL to 5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
Figure 1 / 3
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| Kit Specifications | |
| Sample Type | Plasma/Serum |
| Anti-Coagulant (for Plasma)† | EDTA or Citrate |
| Sample Volume Range | 50 to 200 μL |
| Minimum Elution Volume | 10 μL |
| Maximum Elution Volume | 25 μL |
| Time to Complete 10 Purifications | 15 – 20 minutes |
| Size of RNA Purified | All sizes, including miRNA and small RNA (<200 nt) |
| Average Yields¥ | Variable depending on specimen |
† This kit is suitable for the isolation of RNA from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR
¥ Please check page 5 for Average Plasma/Serum Yields and Common RNA Quantification Methods
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
| Component | Cat. 73700 (50 preps) | Cat. 73710 (20 preps) | Cat. 73720 (10 preps) |
|---|---|---|---|
| Lysis Buffer A | 2 x 20 mL | 100 mL | 1 x 130 mL 1 x 30 mL |
| Wash Solution A | 18 mL* | 1 x 38 mL* 1 x 18 mL* | 38 mL* |
| Elution Solution A | 6 mL | 6 mL | 6 mL |
| Elution Buffer F | – | – | 15 mL |
| EXTRACLean Micro Spin Columns | 50 | – | – |
| EXTRACLean Mini Spin Columns | – | 20 | 10 |
| EXTRACLean Midi Spin Columns | – | 20 | – |
| EXTRACLean Maxi Spin Columns | – | – | 10 |
| Collection Tubes | – | 20 | 10 |
| Elution Tubes (1.7 mL) | 50 | 20 | 10 |
| Product Insert | 1 | 1 | 1 |
Introduction
Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments, <1000bp (DNA). HiPure Circulating DNA Mini Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine. Samples can be fresh or frozen (provided that they have not been frozen and thawed more than once).
Specifications
| Features | Specifications |
| Main Functions | Isolation circulating DNA from 1ml plasma, serum, body fluids |
| Applications | qPCR, liquid or solid chip analysis, hybridization and SNP detection, etc. |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Serum, plasma and other cell-free fluid samples |
| Sample amount | 1ml |
| Elution volume | ≥30μl |
| Time per run | ≤50 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100µg |
This product is based on silica gel purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption plate and filter column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10 Mm Tris,pH 8.0).
Kit Contents
| Contents | D318102 | D318103 |
| Purification Times | 50 Preps | 250 Preps |
| Buffer ACL | 50 ml | 250 ml |
| Buffer ACB* | 60 ml | 300 ml |
| Buffer DCW1* | 22 ml | 88 ml |
| Buffer DCW2* | 10 ml | 50 ml |
| Proteinase K | 120 mg | 540 mg |
| Protease Dissolve Buffer | 10 ml | 30 ml |
| Carrier RNA | 110 μg | 310 μg |
| Nuclease Free Water | 10 ml | 30 ml |
| HiPure CFDNA Mini Columns | 50 | 250 |
| 2 ml Collection Tubes | 100 | 500 |
Storage and Stability
Proteinase K and carrier RNA should be stored at 2–8°C upon arrival. However, short-term storage(up to 12 weeks) at room temperature (15–25°C) does not affect their performance. The remainingkit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under these conditions.The entire kit can be stored at 2–8°C, but in this case buffers shouldbe redissolved before use. Make sure that all buffers are at room temperature when used.
Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments,
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