These kits provide a fast, reliable and convenient spin column method for the isolation of high quality, high purity and inhibitor-free cell-free circulating DNA (cfc-DNA) from plasma/serum sample. These kits are designed to isolate all sizes of cfc-DNA from either fresh or frozen plasma/serum samples and the purified DNA is eluted into a flexible elution volume ranging from 25 µL to 50 µL. The purified plasma/serum cfc-DNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive PCR and Southern Blot analysis, microarrays and NGS.
Background
Plasma/Serum cell-free circulating DNA (cfc-DNA) has the potential to provide biomarkers for certain cancers and disease states as well as fetal DNA in maternal blood. Currently, significant advancements are being made in utilizing cfc-DNA as biomarkers for the early diagnosis, prognosis and monitoring of therapy for several cancer types and autoimmune diseases. Cell-free mitocondrial DNA (cfmtDNA) is also under investigation for its clinical significance. This cfc-DNA is usually present as short fragments of less than 1000 bp. In addition, cell-free fetal DNA has been widely used as a non-invasive method for prenatal diagnosis including early identification of fetal sex, genetic studies for families at high risk for inherited genetic disorders, screening for Rhesus factor, screening for aneuploidy and identification of preeclampsia.
Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit Dx
For serum input volumes 10 µL – 200 µL. Purify high-quality DNA in 15-20 minutes
Plasma/Serum Cell-Free Circulating DNA Purification Mini Kit Dx
For serum input volumes 200 µL – 500 µL. Purify high-quality DNA in 15-20 minutes.
Plasma/Serum Cell-Free Circulating DNA Purification Midi Kit Dx
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 25 µL to 50 µL. All components for the purification & concentration are provided in one convenient & fast kit for the easy processing of large input volumes of bodily fluids. For a schematic workflow of the protocol click here to view. For serum input volumes 1 mL – 4 mL. Purify high-quality DNA in 90 minutes.
Plasma/Serum Cell-Free Circulating DNA Purification Maxi Kit Dx
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 25 µL to 50 µL. All components for the purification & concentration are provided in one convenient & fast kit for the easy processing of large input volumes of bodily fluids. For a schematic workflow of the protocol click here to view. For serum input volumes 5 mL – 10 mL. Two kits in one – purify and concentrate DNA from bodily fluids using a convenient two column system.
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| Kit Specifications | |
| Minimum Plasma/Serum Input | 10 μL |
| Maximum Plasma/Serum Input | 200 μL |
| Size of DNA Purified | ≥ 50 bp |
| Elution Volume | 25-50 μL |
| Time to Complete 10 Purifications | 15-20 minutes |
| Average Yields* | Variable depending on specimen |
*Please check page 7 in the product manual for the Plasma/Serum Average Yields and the Common DNA Quantification Methods.
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature (15-25°C) for up to 2 years without showing any reduction in performance. Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Kits contain ready-to-use Proteinase K solution, which is dissolved in a specially prepared storage buffer. The Proteinase K is stable for up to 2.5 years after delivery when stored at room temperature. To prolong the lifetime of Proteinase K, storage at 2–8°C is recommended.
| Component | Cat. Dx55100 (50 preps) | Cat. Dx55500 (50 preps) | Cat. Dx55600 (20 preps) | Cat. Dx55800 (10 preps) |
|---|---|---|---|---|
| Binding Buffer B | 2 x 40 mL | 40 mL | 2 x 85 mL 1 x 12 mL | 2 x 85 mL 1 x 40 mL |
| Proteinase K | 2 x 1 mL | 0.6 mL | 6.5 mL | 8 mL |
| Solution WN | 18 mL | 18 mL | 18 mL | 18 mL |
| Wash Solution A | 38 mL | 18 mL | 18 mL | 18 mL |
| Elution Buffer B | 8 mL | 8 mL | 30 mL | 30 mL |
| Micro Spin Columns | 50 | 50 | 20 | 10 |
| Mini Spin Columns | 50 | – | – | – |
| Midi Spin Columns | – | – | 20 | – |
| Maxi Spin Columns | – | – | – | 10 |
| Collection Tubes | 100 | 50 | 20 | 10 |
| Elution Tubes (1.7 mL) | 100 | 50 | 20 | 10 |
| Product Insert | 1 | 1 | 1 | 1 |
The Opentrons 24-well Aluminum Block can be placed directly on the OT-2/Opentrons Flex deck or on the Opentrons Temperature Module. This block acts as a 24 position rack to keep 2mL tubes, 1.5mL tubes, and the NEST 0.5mL tubes at constant temperature. The aluminum block holds temperatures between 4°C and 95°C when used with the Opentrons Temperature Module (GEN2).
The Opentrons 24-well Aluminum Block can be placed directly on the OT-2/Opentrons Flex deck or on the Opentrons Temperature Module. This block acts as a 24 position rack to keep 2mL tubes, 1.5mL tubes, and the NEST 0.5mL tubes at constant temperature. The aluminum block holds temperatures between 4°C and 95°C when used with the Opentrons Temperature Module (GEN2).
The series of DNA Size Selection Kits (Magnetic Beads) were developed for DNA size selection using magnetic beads. A total of 11 kits are available, with different selection ranges spanning from 50 bp to over 10 kb. The kits provide a simple and quick approach for the enrichment of a specific range of DNA fragments. The kit workflow allows double-sided or single-sided size selection for specific size cutoffs.
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DNA size selection is a selective capture of DNA fragments of a specific range of size for next-generation sequencing (NGS) library preparations, PCR, ChIP assay, DNA ligations, endonuclease digestions, adapter removal, and other genomics and molecular biology applications. DNA size selection is preferred after NGS library prep in most of the cases. 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 technology uses paramagnetic particles, also known as SPRI (Solid Phase Reversible Immobilization) beads, to bind DNA reversibly and selectively. DNA fragments can be size-selected and purified by changing the properties of the magnetic beads or SPRI beads. The magnetic beads can easily separate the beads-binding DNA from the contaminants and unwanted components in the samples. The samples after DNA size selection are free of contaminants such as buffer components, enzymes, proteins, salts, dNTPs, primers, and adapters. Our proprietary magnetic beads reagents improve yield, selectivity, and reproducibility.
Specific DNA fragments at a certain length range can be purified simply using magnetic separation with different beads components, avoiding tedious and time-consuming gel extraction and column-based purification. The magnetic beads method is popular for common DNA size selection, including library size selection. The first beads-binding step, referred to as the right-side clean-up, removes large DNA fragments. The large DNA fragments are bound to the beads and are discarded. The desired DNA fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-binding, referred to as the left-side clean-up. The double-size selected DNA fragments are eluted after ethanol rinsing.
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A single clean-up is needed for DNA size selection with large fragments. In this case, only the large DNA fragments are bound to the beads. The selected larger DNA fragments are eluted after ethanol rinsing.
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