Plasma/Serum Total cfc-Nucleic Acid Advanced Purification Kit
For rapid and simple combined purification of all sizes of cell-free circulating DNA (cfc-DNA), and circulating tumour DNA (ct-DNA) and circulating/exosomal RNA, including microRNA
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For research use only and NOT intended for in vitro diagnostics.
Plasma/Serum Total cfc-Nucleic Acid Advanced Purification Kit
For rapid and simple combined purification of all sizes of cell-free circulating DNA (cfc-DNA), and circulating tumour DNA (ct-DNA) and circulating/exosomal RNA, including microRNA
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Features and Benefits
- Versatile plasma/serum input ranging from 1 mL to 6 mL
- No phenol extractions
- No carrier RNA
- Minimal high molecular weight gDNA contamination in the purified cfc-DNA
- Bind and elute all RNA irrespective of size or GC content, without bias
- Concentrate circulating RNA and exosomal RNA into a flexible elution volume ranging from 25 µL to 50 µL
- Purify superior-quality and superior-quantity RNA in 45 minutes
- Compatible with fresh, preserved or frozen serum/plasma prepared from blood collected on either Norgen's cf-DNA/cf-RNA Preservative Tubes (Cat. 63950, 63960), Cell-Free DNA BCT ® (Streck), EDTA or Citrate
- Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen's NGS services
- Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
Norgen’s Plasma/Serum Total cfc-Nucleic Acid Advanced Purification Kit provides fast, reliable and simple procedures for isolating the highest quality and quantity of all sizes of circulating DNA and circulating RNA from various amounts of plasma/serum ranging from 1 mL up to 6 mL combined into one elution. Purification is based on using Norgen’s proprietary resin separation matrix. The kits are designed to isolate total cfc-nucleic acid from either fresh, preserved or frozen plasma/serum samples. Moreover, these kits allow the user to elute the purified cfc-RNA into a flexible elution volume ranging from 25 µL to 50 µL. The purified plasma/serum total cfc-nucleic acid is eluted in an Elution Solution that is compatible with all downstream applications including PCR, qPCR, methylation-sensitive PCR and Southern Blot analysis, microarrays and NGS.
Details
Supporting Data
Figure 1. A representative Bioanalyzer RNA Pico Chip trace showing the quality/quantity of the RNA purified from 4 mL K2 EDTA plasma using A) Norgen’s Plasma/Serum Total cfc-Nucleic Acid Advanced Purification Kit (Automated Isolation), B) Norgen’s Plasma/Serum Total cfc-Nucleic Acid Advanced Purification Kit (Manual Isolation), C) Competitor Q's ccfDNA/RNA Kit and D) Competitor Q's Circulating Nucleic Acid Kit. All purification methods showed the correct cf-RNA size on the bioanalyzer trace however the amount of the cf-RNA recovered using Norgen’s technology (as can be noticed by the height of the cf-RNA peak) was significantly higher than that for the cf-RNA purified using the silica-based technology.
Figure 2. A representative Bioanalyzer High Sensitivity DNA Chip trace showing the quality/quantity of the cf-DNA purified from 4 mL K2 EDTA plasma A) Norgen’s Plasma/Serum Total cfc-NA Advanced Purification Kit (Automated Isolation), B) Norgen’s Plasma/Serum Total cfc-NA Advanced Purification Kit (Manual Isolation), C) Competitor Q's ccfDNA/RNA Kit and D) Competitor Q's MinElute ccfDNA Midi Kit. All purification methods showed the correct cf-DNA size (~180bp) on the bioanalyzer trace however the amount of the cf-DNA recovered using Norgen’s technology (as can be noticed by the height of the cf-DNA peak) was significantly higher than recovered using the silica-based technology.
Figure 3. Real-Time PCR amplification of miR-21, miR-16 (cfc-RNA targets) and ALU 247, ALU 115 (cfc-DNA targets) from Total cfc-Nucleic acid purified from 4mL K2EDTA plasma using different purification methods.. Norgen’s SiC technology, manual and automated procedure, was superior as compared to the amplification of the same targets amplified from Total Nucleic aicd purified from K2EDTA plasma using the Silica-based technology represented by Competitor Q.
Kit Specifications
|
|
Minimum Plasma/Serum Input
|
1 mL |
Maximum Plasma/Serum Input
|
6 mL
|
Size of cfc-RNA Purified
|
All sizes, including miRNA and small RNA (< 200 nt)
|
Optimal size of purified cfc-DNA
|
146 - 180 bp
|
Elution Volume |
25-50 µL
|
Time to Complete 10 Purifications |
45 minutes
|
Average Yields* | Variable depending on specimen |
*Please check page 5 in the product manual for the Plasma/Serum Average Yields and the 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. The kit contains a ready-to-use Proteinase K, which is dissolved in a specially prepared storage buffer. The buffered Proteinase K is stable for up to 2 years after the date of shipment when stored at room temperature.
Components | Cat. 68100 (50 preps) |
---|---|
Binding Buffer A | 20 mL |
Proteinase K | 6.5 mL |
Slurry E | 12.5 mL |
Lysis Buffer A | 130 mL |
Wash Solution A | 38 mL |
Elution Buffer C | 2 x 8 mL |
Elution Buffer F | 15 mL |
Elution Solution A | 20 mL |
Micro Spin Columns | 50 |
Mini Filter Spin Columns | 50 |
Collection Tubes | 100 |
Elution Tubes (1.7 mL) | 100 |
Product Insert | 1 |
*Please check page 5 in the product manual for the Plasma/Serum Average Yields and the Common RNA 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 CFC-RNA Advanced Purification Kit 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.