Name: Preserved Blood RNA Purification Kit II (for use with Paxgene Blood RNA Tubes)
Brand: Norgen Biotek Corp.
SKU: 43500
Price: 525.000000 USD
Availability: InStock

Overview

Compatible with PAXgene Blood RNA Tubes
Isolate true total RNA including siRNA and microRNA
No phenol extractions
Purify high-quality RNA in 30 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 Preserved Blood RNA Purification Kit II provides a rapid method for the isolation and purification of total RNA from blood that has been preserved using PAXgene Blood RNA Tubes*. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA). The RNA is preferentially purified from other cellular components such as proteins, without the use of phenol or chloroform.

Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. Norgen’s proprietary resin provides superior affinity to the full size range of RNA molecules, resulting in large and small RNA (miRNA) purification with better linearity and sensitivity. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time RT-PCR, RT-PCR, Northern blotting, RNase protection and primer extension, expression profiling, miRNA cloning and amplification and Next Generation Sequencing.

* PAXgene Blood RNA Tubes are a product of PreAnalytiX GmbH. PAXgene Blood RNA Tubes are not provided.

Details

Supporting Data

Figure 1. High Quality of Isolated RNA with Complete Size Range. Unlike most competitor’s kits, Norgen's Preserved Blood RNA Purification Kit II (for use with Paxgene Blood RNA Tubes) allows for the isolation of all sizes of RNA, from the very large RNA down to the microRNA without the use of phenol. Total RNA was isolated from preserved blood using Norgen's Preserved Blood RNA Purification Kit II (for use with PAXgene Blood RNA Tubes) and a leading competitors kit. The purified RNA was then resolved on a 1.2% formaldehyde-agarose gel. As it can be seen, only Norgen's kit was able to isolate the full diversity of RNA species, including all the small RNA species.

Figure 2. High Yield of a Diversity of RNA Species. Norgen's Preserved Blood RNA Purification Kit II (for use with PAXgene Blood RNA Tubes) effectively recovers all sizes of RNA from large mRNA to small RNA, including microRNAs. Total RNA was isolated from human blood collected in a PAXgene Blood RNA Tube followed by RNA isolation using Norgen's Preserved Blood RNA Purification Kit. II The purified RNA was then used as the template in a RT-qPCR for detecting the S15 gene (blue lines, in duplicate) and for detecting miR-21 (red lines, in duplicate). As it can be seen, Norgen's kit isolated complete range of RNA including high yields of mRNA and microRNA.

Kit Specifications
Maximum Column Binding Capacity	50 μg
Maximum Column Loading Volume	650 μL
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Time to Complete 10 Purifications	30 minutes
Average Yield	5 - 20 μg per 2.5 mL preserved human blood

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 date of shipment. The DNase I should be stored at -20°C.

Component	Cat. 43500 (50 preps)
NPX1	2 x 110
NPX2	40 mL
NPX3	22 mL
NPX4	6 mL
NPX5	6 mL
DNase I	1 vial
Mini Spin Columns	50
Collection Tubes	50
Elution Tubes (1.7 mL)	50
Product Insert	1

Documentation

PROTOCOLS

(43500) Preserved Blood RNA Purification Kit II - Protocol (50 prep)

SAFETY DATA SHEETS

43500 - Preserved Blood RNA Purification Kit II (for use with PAXgene Blood RNA Tubes) - SDS

FLYERS

Flyer

RELATED BLOGS

FAQs

Spin Column

Why do I have poor RNA recovery?

Poor RNA recovery could be due to one or more of the following:

Incomplete lysis of blood.
Ensure that the appropriate amount of NPX2 was used.
An alternative elution solution was used.
It is recommended that the NPX5 Solution supplied with this kit be used for maximum RNA recovery.
Ethanol was not added to the lysate.
Ensure that the appropriate amount of ethanol is added to the lysate before binding to the column.
Ethanol was not added to NPX3.
Ensure that 50 mL of 96-100% ethanol is added to the supplied NXP3 prior to use.
Insufficient centrifugation time and speed when pelleting the stabilized blood RNA before addition of NPX2 for Lysis.
Centrifuge the PAXgene™ Tube for 10 minutes at 3,000–5,000 x g.
Low RNA Content in Sample.
Amount of cellular RNA could vary drastically in different individuals. In particular, blood from healthy individuals with low white blood cell count is expected to have lower RNA yield

I noticed the columns are getting clogged. What causes this?

Column clogging can result from one or combination of the following factors:

Insufficient solubilization of blood.
Ensure that the appropriate amount of NPX1 and NPX2 was used.
Centrifuge temperature is too low.
Ensure that the centrifuge remains at room temperature throughout the procedure. Temperatures below 15°C may cause precipitates to form that can cause the columns to clog.

Why is the RNA degrading?

RNA can be degraded due to following factors:

RNase contamination.
RNases may be introduced during the use of the kit. Ensure proper procedures are followed when working with RNA. Please refer to “Working with RNA” at the beginning of the protocol's user guide.
Procedure not performed quickly enough.
In order to maintain the integrity of the RNA, it is important that the procedure be performed quickly.
Improper storage of the purified RNA.
For short term storage RNA samples may be stored at -20°C for a few days. It is recommended that samples be stored at -70°C for longer term storage.

Why is the RNA not performing well in downstream applications?

If the RNA does not perform well in downstream applications, It may be due to one or more of the following:

RNA was not washed 3 times with NPX3.
Traces of salt from the binding step may remain in the sample if the column is not washed 3 times with NXP3. Salt may interfere with downstream applications, and thus must be washed from the column.
Ethanol carryover.
Ensure that the dry spin under the Column Wash procedure is performed, in order to remove traces of ethanol prior to elution. Ethanol is known to interfere with many downstream applications.

I noticed genomic DNA is contaminated. Why did this happen?

Genomic DNA contamination could occur if large amount of starting material used. Perform the DNase I digestion on the RNA sample prior to elution using the provided DNase I solution and NPX4 solution.

Citations

Title	The Development of Translational Biomarkers as a Tool for Improving the Understanding, Diagnosis and Treatment of Chronic Neuropathic Pain
Journal	Molecular Neurobiology. 2017
Authors	Buckley DA, Jennings EM, Burke NN, Roche M, McInerney V, Wren JD, Finn DP, McHugh PC.

Title	Evaluation of Existing Methods for Human Blood mRNA Isolation and Analysis for Large Studies
Journal	PLoS One. 2016.
Authors	Meyer A, Paroni F, Günther K, Dharmadhikari G, Ahrens W, Kelm S, Maedler K

Preserved Blood RNA Purification Kit II (for use with PAXgene Blood RNA Tubes)