Name: Plasma/Serum cfc-DNA/cfc-RNA Advanced Fractionation Kit
Brand: Norgen Biotek Corp.
SKU: 68300
Price: 1079.000000 USD
Availability: InStock

Features and Benefits

Versatile plasma/serum input ranging from 1 mL to 6 mL
No phenol extractions or 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
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 cfc-DNA/cfc-RNA Advanced Fractionation Kit provides fast, reliable and simple procedures for the fractionation of 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 into two separate elutions. 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 and cfc-DNA 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. Representative Bioanalyzer RNA Pico Chip traces and High Sensitivity DNA Chip traces showing the quality/quantity of the RNA fraction and the DNA fraction purified simultaneously from the same 4 mL K2 EDTA plasma using both automated and manual isolation. A) Norgen’s Plasma/Serum cfc-DNA/cfc-RNA Advanced Fractionation Kit (DNA Fraction from Automated Isolation), B) Norgen’s Plasma/Serum cfc-DNA/cfc-RNA Advanced Fractionation Kit (DNA Fraction from Manual Isolation), C) Norgen’s Plasma/Serum cfc-DNA/cfc-RNA Advanced Fractionation Kit (RNA Fraction from Automated Isolation), D) Norgen’s Plasma/Serum cfc-DNA/cfc-RNA Advanced Fractionation Kit RNA Fraction from Manual Isolation). All purification methods showed the correct cf-RNA size on the Bioanalyzer trace.

Figure 2. RT-qPCR amplification of miR-21 and miR-16 from the RNA fraction purified from 4mL K2EDTA plasma using Norgen’s Plasma/Serum cfc-DNA/cfc-RNA Advanced Fractionation Kit, both manually and automated. There is no difference in the amplification of both miR-21 and miR-16 between the automated and the manual isolation.

Figure 3. Real-Time PCR amplification of ALU 247 and ALU 115 from the DNA fraction purified from 4mL K2EDTA plasma using Norgen’s Plasma/Serum cfc-DNA/cfc-RNA Advanced Fractionation Kit, both manually and automated. There is no difference in the amplification of both ALU 247 and ALU 115 between the automated and the manual isolation.

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.

Component	Cat. 68300 (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	2 x 38 mL
Elution Buffer C	2 x 8 mL
Elution Buffer F	1 x 6 mL 1 x 15 mL
Elution Solution A	6 mL
Micro Spin Columns	100
Mini Filter Spin Columns	50
Collection Tubes	150
Elution Tubes (1.7 mL)	150
Product Insert	1

Documentation

PROTOCOLS

(68300) Plasma/Serum cfc-DNA/cfc-RNA Advanced Fractionation Kit - Protocol (50 Preps)

SAFETY DATA SHEETS

68300 - Plasma/Serum cfc-RNA/cfc-DNA Advanced Fractionation Kit - SDS

RELATED BLOGS

FAQs

Spin Column

What if a variable speed centrifuge is not available and the speed differs from the recommended?

A fixed speed centrifuge can be used, however reduced yields may be observed.

At what temperature should I centrifuge my samples?

All centrifugation steps are performed at room temperature. Centrifugation at 4°C will not adversely affect kit performance.

What if I added more or less of the specified reagents' volume?

Adding more or less than the specified volumes may reduce both the quality and the quantity of the purified RNA. Eluting your Nucleic Acid in high volumes will increase the yield but will lower the concentration. Eluting in small volumes will increase the concentration but will lower the overall yield.

What If I forgot to do a dry spin before my final elution step?

Your purified Nucleic Acid will be contaminated with the Wash Solution A. This may reduce the quality of your purified Nucleic Acid and will interfere with your downstream applications.

Can I perform a second elution?

Yes, but it is recommended that the 2nd elution be in a smaller volume (50% of 1st Elution). It is also recommended to perform the 2nd elution into a separate elution tube to avoid diluting the 1st elution.

What if my incubation time varied from what is specified in the product manual?

Varying the incubation time will result in a reduction in your Nucleic Acid yields.

Why do my samples show very low RNA yield?

Plasma/Serum samples contain very little cfc-Nucleic Acid. This varies from individual to individual. In order to increase the yield, the amount of Plasma/Serum input could be increased.

Why does my purified cfc-Nucleic Acid not perform well in downstream applications?

If a different Elution solution was used other than the one provided in the kit, the buffer should be checked for any components that may interfere with the application. Common components that are known to interfere are high salts (including EDTA), detergents and other denaturants. Check the compatibility of your Elution Buffer with the intended use.

Citations

Title	EP04.01-04 Detecting KRAS Mutations in Peripheral Blood Extracellular Vesicles in Non-Small Cell Lung Cancer Using Digital Droplet PCREP04.01-04 Detecting KRAS Mutations in Peripheral Blood Extracellular Vesicles in Non-Small Cell Lung Cancer Using Digital Droplet PCR EP04.01-04 Detecting KRAS Mutations in Peripheral Blood Extracellular Vesicles in Non-Small Cell Lung Cancer Using Di
Citation	Journal of thoracic oncology 2023.
Authors	C.J. Zahra, K. Chee, I.A. Yang and K.M. Fong

Plasma/Serum cfc-DNA/cfc-RNA Advanced Fractionation Kit